CN106279200A - There is the Diterpene compounds of hypolipidemic activity, Preparation Method And The Use - Google Patents

Abstract

The present invention provides a class of abietane-type diterpenoids with blood lipid-lowering activity, its preparation method and application, specifically, the compound has the structure shown in formula I, wherein the definition of each group is as in the description described in . The compound of the present invention has the activity of increasing low-density lipoprotein uptake in vivo or in vitro, and has the potential of being a new type of blood lipid-lowering drug.

Description

Abietin-type diterpenoid compound with hypolipidemic activity, preparation method and use thereof

technical field

The invention belongs to the field of medicinal chemistry, and more specifically relates to a new class of abietane-type diterpenoids with blood lipid-lowering activity, its preparation method and its use in drugs for reducing blood fat and atherosclerosis.

Background technique

Atherosclerosis (AS) refers to the deposition of lipids (mainly cholesterol and cholesteryl lipid) in the intima and subintimal of the arterial wall of the artery and its branches, accompanied by the migration of smooth muscle cells in the middle layer to the subintimal hyperplasia, Yellow or gray-yellow plaques like atheroma are formed, which thicken and harden the arterial wall, lose its elasticity, and make the lumen smaller. AS is the most common and important lesion in arteriosclerotic vascular diseases, and has been the main cause of death in western developed countries. In recent years, with the improvement of Chinese people's living standards and changes in eating habits, the disease has also become the main cause of death in my country, and the incidence rate among people over 60 years old in China is as high as 79.9%.

A large number of research data show that hyperlipidemia is the main risk factor for human atherosclerotic diseases. The direct damage of high blood cholesterol (TC) and/or triglyceride (TG) or low high-density lipoprotein cholesterol (HDL-C) (called dyslipidemia in modern medicine) can accelerate systemic atherosclerosis, Because vital organs of the whole body rely on arteries for blood and oxygen supply, once the arteries are blocked by atheromatous plaques, serious consequences will result.

Cholesterol plays an important role in metabolic syndrome and atherosclerosis. 70% of cholesterol in the blood is carried by low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL), and 90% of LDL is carried by low-density lipoprotein Protein receptor (LDLR) pathway for clearance. LDLR is the main receptor for liver recycling and uptake of LDL. It plays an important role in the lipoprotein metabolism of steroid-derived tissues such as the liver and adrenal cortex, testis, and ovary. Its main function is to participate in the metabolism of LDL. LDLR gene mutation is one of the main causes of hereditary hyperlipidemia.

Lowering blood lipids is the most effective treatment for atherosclerosis, and statins are currently the most widely used and most powerful lipid-lowering drugs in clinical practice. As a star drug for lowering blood lipids on the market, it can competitively inhibit the activity of the rate-limiting enzyme hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase in the process of cholesterol synthesis in the body, reducing free cholesterol in cells, and through feedback regulation Function, up-regulate the number of low-density lipoprotein (LDL) receptors on the cell surface, accelerate the catabolism of LDL in plasma, and then regulate the body's blood lipid level. With the widespread use of statins, problems such as adverse reactions and interactions with other drugs have gradually attracted the attention of clinical medical staff, especially after the Bistin incident has aroused concerns about the safety of statins blood lipid-regulating drugs. of great concern. The adverse reactions of statins that have been reported so far include liver toxicity, muscle toxicity (rhabdomyolysis), elevated blood sugar levels, osteoarthritis, and joint pain. Statins are not effective for all patients with hyperlipidemia and high risk of atherosclerosis. Clinical studies have shown that the use of pravastatin (pravastatin) or atorvastatin (atorvastatin) intensive treatment of coronary heart disease patients after an average of 24 months , LDL cholesterol levels have been reduced to varying degrees, but there is still a high incidence of cardiovascular disease. In addition, a large proportion of patients are insensitive to statins and cannot rely on statins to prevent the development of atherosclerosis. Therefore, the task of developing new lipid-lowering drugs is imminent.

In summary, there is an urgent need in this field to provide new compounds with hypolipidemic activity.

Contents of the invention

The object of the present invention is to provide a class of compounds with hypolipidemic activity.

The first aspect of the present invention provides a use of a compound represented by the following formula I, its dimer, or a pharmaceutically acceptable salt thereof:

in,

Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl;

X is selected from the following group: CHRe, NRe, O, C=Re, C-Re, N; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl;

M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl; or Re, Rf are each independently a substituent selected from the following group:

Or Re and Rf jointly constitute a structure selected from the following group: -O-, -C ₁ -C ₆ alkylene -, -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene -OC ₁ -C ₆ alkylene -,

Or one of Re, Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; Wherein, the substitution means that one or more hydrogen atoms on the group are replaced by =O;

Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl;

Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or Unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group: halogen, -OH, -COOH;

Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-;

Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH;

Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or form a 5-9 membered epoxy ring or lactone ring together with Rc;

R" is selected from the group consisting of H, -COOH, -OH, -Me;

Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group;

Dotted lines are single or double bonds;

and It is a substituted or unsubstituted aromatic ring (benzene ring or benzoquinone);

It is characterized in that the compound is used for one or more purposes selected from the following group:

a) preparing a pharmaceutical composition for lowering blood fat;

b) preparing a pharmaceutical composition for lowering low-density lipoprotein (LDL) levels;

c) preparing a pharmaceutical composition for stabilizing LDLR gene stability;

d) preparing a pharmaceutical composition for up-regulating LDLR gene expression;

e) preparing a pharmaceutical composition for increasing the number of LDLR receptors on the surface of liver cells;

f) preparing a pharmaceutical composition for reducing degradation of LDLR receptors;

g) preparing a pharmaceutical composition for reducing the concentration of TC and/or TG in blood;

h) preparing a pharmaceutical composition for increasing the concentration of high-density lipoprotein (HDL) in blood;

i) preparing a pharmaceutical composition for improving liver function damage.

In another preferred example, the dimer of the compound of formula I has the structure shown in the following formula Ia:

In the formula,

Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl;

The definitions of the remaining groups are as described in the first aspect of the present invention.

In another preferred example, X is selected from the following group: CHRe, O, C=Re, C-Re; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl;

M is selected from the following group: CHRf, O, C=Rf, C-Rf; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ - C ₆ alkyl, -OC ₁ -C ₆ acyl;

Or Re and Rf jointly constitute a structure selected from the following group: -O-, -C ₁ -C ₆ alkylene -, -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene -OC ₁ -C ₆ alkylene -;

Or one of Re and Rf together with Ri constitutes a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; wherein, the Substitution means that one or more hydrogen atoms on the group are replaced by =O;

Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl;

Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted Or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution refers to one of the groups or multiple hydrogen atoms are substituted by substituents selected from the following group: halogen, -OH, -COOH;

Ri is selected from the group consisting of H, -COOH, -Me; or Ra and Ri jointly form -C(O)-O-;

The definitions of the other groups are as described above.

In another preferred embodiment, Rc is H.

In another preferred embodiment, Ra, Rb, Rc, and Rd are each independently selected from the following group: H, -OH, -OMe;

X is selected from the following group: CHRe, O, C=Re, C-Re; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, =O;

M is selected from the following group: CHRf, O, C=Rf, C-Rf; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, =O;

And X and M are not O at the same time;

Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₃ alkyl, -OH; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by Substituents selected from the group consisting of: halogen, -OH, -COOH;

Rj is H.

In another preferred example, the compound of formula I is an abietane-type diterpenoid.

In another preferred example, the compound of formula I is a plant extract selected from the group consisting of cedar bark extract, Pork cedar bark extract, Pork cedar fruit extract, Chinese fir bark extract, Branosic acid, carnosol.

In another preferred example, the pharmaceutical composition is also used for purposes selected from the following group:

j) (in vitro non-therapeutic) increasing the uptake rate of LDL by hepatocytes;

k) Up-regulation (non-therapeutically) of the number of LDLR receptors on the surface of hepatocytes in vitro.

In another preferred example, in the pharmaceutical composition, the effective dose of the compound of formula I is 0.1-50 mg/kg body weight, preferably 1-20 mg/kg body weight.

In another preferred example, the pharmaceutical composition is a dosage form selected from the group consisting of oral dosage form and injection dosage form.

The second aspect of the present invention provides a kit, characterized in that the kit contains:

(i) a first container, and the active ingredient (a) compound of formula I contained in the first container; or the medicine containing the active ingredient (a);

(ii) a second container, and the active ingredient (b) statin drug, or a pharmaceutically acceptable salt thereof; or a drug containing active ingredient (b) contained in the second container; and

(iii) an instruction sheet describing an explanation for reducing the low-density lipoprotein content in the body of a user by co-administering the active ingredient (a) and the active ingredient (b).

The third aspect of the present invention provides a compound represented by the following formula I, or a dimer thereof:

in,

Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl;

X is selected from the group consisting of CHRe, NRe, O, C=Re, C-Re, N;

M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, said Re, Rf are each independently a substituent selected from the following group:

Or Re and Rf jointly constitute a structure selected from the group consisting of: -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene - OC ₁ -C ₆ alkylene-,

Or one of Re, Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; Wherein, the substitution means that one or more hydrogen atoms on the group are replaced by =O;

Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl;

Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or Unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group: halogen, -OH, -COOH;

Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-;

Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH;

Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or together with Rc form a 5-9 membered oxygen-containing ring (preferably an epoxy ring or a lactone ring);

R" is selected from the group consisting of H, -COOH, -OH, -Me;

Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group;

Dotted lines are single or double bonds;

and It is a substituted or unsubstituted aromatic ring (benzene ring or quinone).

The fourth aspect of the present invention provides a compound represented by the following formula I, or a dimer thereof:

in,

Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl;

X is O;

M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl;

Or said Rf is a substituent selected from the following group:

Or one of Rf and Ri or Rg together constitutes a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; wherein, the Substitution means that one or more hydrogen atoms on the group are replaced by =O;

Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or Unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group: halogen, -OH, -COOH;

Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-;

Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH;

Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or together with Rc form a 5-9 membered oxygen-containing ring (preferably an epoxy ring or a lactone ring);

R" is selected from the group consisting of H, -COOH, -OH, -Me;

Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group;

Dotted lines are single or double bonds;

and It is a substituted or unsubstituted aromatic ring (benzene ring or quinone).

The fifth aspect of the present invention provides a compound represented by the following formula I, or a dimer thereof:

in,

Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl;

X is selected from the following group: CHRe, NRe, O, C=Re, C-Re, N; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl;

M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl;

Or described Re, Rf are each independently a substituent selected from the following group:

Or Re and Rf jointly constitute a structure selected from the group consisting of: -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene - OC ₁ -C ₆ alkylene-,

Or one of Re, Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; Wherein, the substitution means that one or more hydrogen atoms on the group are replaced by =O;

Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl;

Rg is selected from the group consisting of H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or unsubstituted C ₁ -C ₆ alkoxy; wherein, the substitution means that one or more hydrogen atoms on the group are substituted by a substituent selected from the following group: halogen, -OH, -COOH;

Rh is a water solubility improving group, preferably selected from the group consisting of -COOH, -OH, substituted or unsubstituted C ₁ -C ₆ alkylene-OH, substituted or unsubstituted C ₁ -C ₆ alkylene- COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl , -COO-substituted or unsubstituted C ₁ -C ₆ alkoxy; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by substituents selected from the group consisting of halogen, -OH, -COOH;

Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-;

Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH;

Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or together with Rc form a 5-9 membered oxygen-containing ring (preferably an epoxy ring or a lactone ring);

R" is selected from the group consisting of H, -COOH, -OH, -Me;

Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group;

Dotted lines are single or double bonds;

and It is a substituted or unsubstituted aromatic ring (benzene ring or quinone).

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, we will not repeat them here.

Description of drawings

The effect of compounds LSP-6-1～LSP-6-6 in Example 2 on the uptake of low-density lipoprotein in hepatocytes;

The effect of compound LSP-6-6 on the uptake of hepatocyte low-density lipoprotein under different concentrations in Fig. 2 embodiment 2;

Compound up-regulates low-density lipoprotein receptor (LDLR) expression level test result in Fig. 3 embodiment 3;

The influence of Fig. 4 compound LSP-6-6 oral administration on hyperfat model golden hamster total cholesterol level; A among Fig. 4: TC (serum total cholesterol) level, B among Fig. 4: TG (triglyceride) level, C in Figure 4: LDL level; *P<0.05, **P<0.01vs normal control group, #P<0.05, ##P<0.01vs high-fat control group.

detailed description

After long-term and in-depth research, the inventors unexpectedly found that abietane-type diterpenoids have good blood-lipid-lowering activity, and the activity is better than that of existing general-purpose blood-lipid-lowering compounds statins. The compound can reduce the concentration of total cholesterol, triglyceride and low-density lipoprotein and increase the concentration of high-density lipoprotein in vivo or in vitro in a time-dependent and dose-dependent manner. Based on the above findings, the inventors have accomplished the present invention.

the term

As used herein, the term "C ₁ -C ₆ alkyl" or "C ₁ -C ₁₀ alkyl" refers to a straight or branched chain alkyl having 1 to 6 or 1 to 10 carbon atoms, such as methyl, Ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.

The term "C ₃ -C ₆ cycloalkyl" refers to a cycloalkyl group having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, methylcyclobutyl, cyclopentyl, or the like.

As used herein, the term "C ₁ -C ₁₀ acyl" or "C ₁ -C ₄ acyl" refers to a straight or branched chain alkyl/cycloalkyl having 0 to 9 or 0 to 3 carbon atoms /Aryl-carbonyl"structure substituents, such as acetyl, propionyl, butyryl, or similar groups.

The term "C ₂ -C ₁₀ ester group" refers to a substituent in the form of a "straight chain or branched chain alkyl/cycloalkyl/aryl-carbonyl-oxygen atom with 1 to 9 carbon atoms", such as acetyl , propionyl, butyryl, or similar groups.

The term "C ₁ -C ₄ alkylene" refers to a group formed after a C1-C4 alkyl group as described above loses a hydrogen atom, such as -CH ₂ -, -CH ₂ -CH ₂ -, or similar groups .

The term "halogen" refers to F, Cl, Br and I.

The term "C ₆ -C ₁₀ aryl" refers to an aryl group having 6-10 carbon atoms, such as phenyl, naphthyl and the like.

The term "C ₁ -C ₁₀ heteroaryl" refers to a heteroaryl group having 1-10 carbon atoms and one or more heteroatoms selected from O, S and/or N.

In the present invention, the term "comprises", "comprises" or "comprises" means that various components can be applied together in the mixture or composition of the present invention. Accordingly, the terms "consisting essentially of" and "consisting of" are included in the term "comprising".

In the present invention, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for human and/or animal without undue adverse side effects (such as toxicity, irritation and allergic reaction), ie, has a reasonable benefit/risk ratio.

In the present invention, the term "effective amount" refers to the amount of a therapeutic agent that treats, alleviates or prevents a target disease or condition, or exhibits a detectable therapeutic or preventive effect. The precise effective amount for a subject will depend on the size and health of the subject, the nature and extent of the disorder, and the therapeutic agents and/or combination of therapeutic agents chosen for administration. Therefore, it is not useful to prespecify an exact effective amount. However, the effective amount can be determined by routine experimentation, within the judgment of the clinician, for a given situation.

In this article, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on the group are replaced by substituents selected from the following group: halogen, unsubstituted or halogenated C1-C6 alkyl, unsubstituted Substituted or halogenated C2 _{- C6} acyl, unsubstituted or halogenated C1-C6 alkyl-hydroxyl.

Unless otherwise specified, in the present invention, all appearing compounds are intended to include all possible optical isomers, such as single chiral compounds, or mixtures (ie racemates) of various chiral compounds. In all compounds of the present invention, each chiral carbon atom may optionally be in R configuration or S configuration, or a mixture of R and S configurations.

As used herein, the term "compound of the present invention" refers to a compound represented by formula I. The term also includes the various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula I.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or a base which is suitable for use as a medicine. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Acids suitable for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, Maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenemethanesulfonic acid, benzenesulfonic acid and other organic acids; and acidic amino acids such as aspartic acid and glutamic acid.

As used herein, the term "dimer" refers to the abietane-type diterpene itself or with other abietane-type diterpenes (including A, B ring-opened or ring-opened abietane-type diterpenes) through C-O-C, Either C-C, or a dimer connected by C-O-C and C-C at the same time.

abietane-type diterpenoids

As a defensive component in plants, abietane-type diterpenoids exist in large quantities in nature and are widely distributed, mainly in higher plants such as Taxaceae, Pinaceae, and Cupressaceae. It is currently known in the art that such diterpenes have antiviral, antitumor, antiulcer, antibacterial, antimalarial and other activities. However, the inventors unexpectedly discovered during the natural product activity screening process that the results of the in vitro HepG2 cell LDL phagocytosis test showed that some abietane-type diterpenes had obvious hypolipidemic activity at a concentration of 5 μM, and the compound LSP-6-6 showed Significant dose-dependent hypolipidemic activity. Compared with the positive control berberine, at the doses of 10 μM and 20 μM, the activity of the two is equivalent. In the animal model, the compound LSP-6-6 was administered intraperitoneally at 30 mg/kg/day, and the total cholesterol (TC), total triglyceride (TG) and LDL levels in the blood of golden hamsters fed with high-fat diet decreased respectively after 20 days 75%, 72% and 58% (relative to the high-fat feeding model group), and no obvious adverse reactions were observed.

As the most preferred embodiment of the present invention, the abietane-type diterpenoid has a structure selected from the following group:

In another preferred embodiment of the present invention, the abietane-type diterpenoid has a structure selected from the following group:

In other embodiments of the present invention, the abietane-type diterpenoids have a structure selected from the following group:

Abiesane-type diterpenes in Cryptomeria (bark):

Abiesane-type diterpenes (*new compounds) from the bark of Pond cedar:

Abiesane-type diterpenes in the fruit of Pondus chinensis:

Abiesane-type diterpenes in fir bark:

Derivatives based on carnosic acid (CA-1):

Pharmaceutical compositions and methods of administration

Since the compound of the present invention has excellent activity of reducing LDL content in blood, the compound of the present invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and compounds containing the present invention are the main The pharmaceutical composition of active ingredients can be used for treating, preventing and alleviating diseases caused by excessive LDL content in blood, such as fatty liver and the like.

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof within a safe and effective amount range and a pharmaceutically acceptable excipient or carrier. Wherein, "safe and effective dose" refers to: the amount of the compound is sufficient to obviously improve the condition without causing severe side effects. Usually, the pharmaceutical composition contains 1-3000 mg (active dose range 3-30 mg/kg) of the compound of the present invention per dose, more preferably 10-2000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use, and must have sufficient purity and low enough toxicity. "Compatibility" herein means that the components of the composition can be blended with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Examples of pharmaceutically acceptable carrier parts include cellulose and derivatives thereof (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as spit ), wetting agent (such as sodium lauryl sulfate), coloring agent, flavoring agent, stabilizer, antioxidant, preservative, pyrogen-free water, etc.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration .

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or extenders, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, For example, glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow agents, such as paraffin; (f) Absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glyceryl monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, hard Calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and others well known in the art. They may contain opacifying agents and, in such compositions, the release of the active compound or compounds may be in a certain part of the alimentary canal in a delayed manner. Examples of usable embedding components are polymeric substances and waxy substances. The active compounds can also be in microencapsulated form, if desired, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

Besides such inert diluents, the compositions can also contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for topical administration of a compound of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required, if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage when administered, for a person with a body weight of 60kg, the daily The dosage is usually 1-2000 mg, preferably 6-600 mg. Of course, factors such as the route of administration and the health status of the patient should also be considered for the specific dosage, which are within the skill of skilled physicians.

Compared with the prior art, the main advantages of the present invention include:

1. The present invention finds for the first time that abietane-type diterpenoids have good blood lipid-lowering activity; the in vitro activity is equivalent to that of berberine at the same concentration; and the activity of some compounds is much higher than that of cypress lactone at a concentration of 5 μM;

2. It was found that the hypolipidemic activity of abietane-type diterpenoids has obvious dose-dependence;

3. Discovered for the first time the blood lipid-lowering mechanism of abietane-type diterpenoids, that is, by up-regulating the number of LDLR receptors on the surface of liver cells to achieve the purpose of clearing LDL;

4. It was found that abietane-type diterpenoids exhibited hypolipidemic activity in vitro and in vivo, and the hypolipidemic effect was shown at a dose as low as 30 mg/kg, which was far lower than that reported by pravastatin and berberis The starting dose of base drugs;

5. In addition to reducing LDL, the abietane-type diterpenoids provided by the present invention can also reduce the concentrations of TC and TG in blood.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For the experimental methods without specific conditions indicated in the following examples, the conventional conditions or the conditions suggested by the manufacturer are usually followed. Percentages and parts are by weight unless otherwise indicated.

In each embodiment, the structure of comparative compound ZBM30 (Nagilactone B (7)) is as follows:

1. Instruments and equipment

In addition, spectrum UV: SHIMADIU UV-2550 and Beckman DU-7 ultraviolet-visible spectrometer;

Infrared spectrum IR: Perkin-Elmer 577 infrared spectrophotometer;

Low resolution mass spectrometry LR-EIMS: Finnigan MAT-95;

High resolution mass spectrometry HR-EIMS: Kratos 1H spectrometer;

Nuclear magnetic resonance spectrum: Varian INOVA 600 nuclear magnetic resonance instrument, Bruker AM-500, AM-400, AM-300 nuclear magnetic resonance instrument, δ (ppm), with TMS as internal standard;

LC-MS: Agilent 1100 liquid phase coupled Bruker esquire mass spectrometer;

Analytical HPLC: Waters 2690Separate Model, Waters PDA 996 detector coupled to Alltch ELSD2000 detector, Millennium 2000 operating system, Waters RP18column (5.0×125mm, 5μm, Waters), flow rate 1.0ml/min, CH3CN (Merck, Germany), H2O (Robust); Jasco HPLC (Chiralcel IA column, 5m, 150×4.6mm), flow rate 0.6ml/min, hexane/ethanol (7:3).

HPLC-MS: Waters 2695Separate Model, Waters PDA 2998 detector coupled to Alltch ELSD2424 detector, 3100Ms detector, SunFireTM C-18column (4.6×100mm, 3.5μm, Waters), flow rate 1.0ml/min, CH3CN (Merck, Germany) , H2O (Robust); Jasco HPLC (Chiralcel IA column, 5m, 150×4.6mm), flow rate 0.6ml/min, hexane/ethanol (7:3).

Preparative HPLC: Varian SD1instrument, Varians 320 single-wavelength detector, C18column (220×25nm, 10μm, Waters), flow rate 15ml/min, CH3CN (Merck, Germany), H2O (Robust);

SpectraMax M2 ^e multifunctional microplate reader (Molecular Instruments, Inc.);

Electrophoresis apparatus and semi-dry electroblotting tank (Bio-Rad Laboratories, Hercules, CA);

PCR machine (Bio-Rad Laboratories, Hercules, CA);

Desktop refrigerated centrifuge (German Hettich company);

752C UV-Vis Spectrophotometer (Shanghai Third Analytical Instrument);

DK-8B electric heating constant temperature water tank (Shanghai Jinghong Experimental Equipment Co., Ltd.);

REVCO carbon dioxide incubator (US REVCO company);

2. Reagents and materials

Column chromatography silica gel: 100-200 mesh, 200-300 mesh silica gel and silica gel H are all produced by Qingdao Ocean Chemical Factory;

TLC thin layer preparation plate: HSGF254 was produced by Yantai Chemical Plant.

MCI resin: CHP20P (75-150μm) is produced by Mitsubishi;

Sephadex LH-20 Sephadex: Pharmacia Biotech AB, Uppsala, Sweden.

Chromogen: 10% sulfuric acid-vanillin solution, iodine;

Plant materials: Cedar cedar, Pond cedar, and Chinese fir were collected in Shexian County, Anhui Province, and Suining, Hunan Province, respectively. They were collected and identified by Associate Professor Shen Jingui from the Herbarium of Shanghai Institute of Materia Medica, and the specimens were preserved in the Herbarium of the Institute.

DMEM and fetal bovine serum (fetal bovine serum, FBS) were purchased from Gibco-BRL (Grand Island, NY); DiI was purchased from biotuim (Hayward, CA); β-actin antibody was purchased from Cell Signaling Technology (Beverly, MA); LDLR Antibodies were purchased from Abcam (Cambridge, United Kingdom); protease inhibitor cocktail was purchased from Calbiochem (San Diego, CA); Immobilon-P transfer membrane (PVDF) was purchased from Millipore Corporation (Bedford, MA); enhanced chemiluminescence reagents (Enhanced chemiluminescence reagents, ECL) were purchased from Pierce (Rockford, IL); medical X-ray film was purchased from China Beyontian Company; golden hamster high-fat diet (0.12% cholesterol, 10% coconut oil) was purchased from Shanghai Slex Company. Protein assay kits were purchased from China Beyontian Company; liver tissue triglyceride TG and total cholesterol TC content assay kits were purchased from Shanghai Rongsheng Biotechnology Company. Blood index (total cholesterol TC, triglyceride TG, high-density lipoprotein HDL, low-density lipoprotein LDL, alanine aminotransferase ALT, aspartate aminotransferase AST) assay kits were purchased from Sichuan Mike Biological Company . Other reagents were purchased from Shanghai Sinopharm Group unless otherwise specified.

The experimental animals were male Syrian golden hamsters, weighing 100±10 g, purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences. Animals were placed in the SPF animal room of Shanghai Institute of Materia Medica, Chinese Academy of Sciences (temperature 23±1°C; humidity 60%), with a natural day/night cycle, and fed with standard food and water. (Experimental approval number: SIMM-AE-2010-10-WYP-01)

The extraction and separation method of embodiment 1 compound LS 41-76

Dry 9.5Kg of cedar bark in the shade and then powder it, soak it in 60L of acetone at room temperature for 20 days, repeat 3 times, combine the extracts, concentrate under reduced pressure to obtain 860g of total extract. The total extract was suspended in 5L of water, extracted with petroleum ether, dichloromethane, and ethyl acetate in sequence, and the organic solvent was evaporated under reduced pressure to obtain 500 g of oil-filled petroleum ether, 120 g of dichloromethane, and 160 g of ethyl acetate. Take 120g of dichloromethane and mix it with 100-200 mesh silica gel (240g) to mix the sample, 200-300 mesh silica gel (2kg) to hold it, and use petroleum ether: acetone system 100:2, 100:5, 10:1, 5:1 , 3:1, 2:1, 1:1, eluted with acetone, combined to obtain 10 fractions after TLC detection. Fraction 2 was subjected to silica gel column chromatography (petroleum ether: ethyl acetate 10:1 ~ 2:1) and Sephadex LH-20 (chloroform-methanol = 6:4) column chromatography to obtain compounds LS-61 (10 mg) and LS -75 (65mg). Fraction 3 was subjected to silica gel column chromatography (petroleum ether: ethyl acetate 8:1 ~ 1:1) and gel Sephadex LH-20 column chromatography (chloroform-methanol = 1:1) to obtain compound LS-41 (68mg) , 43(23mg), 51(40mg), 53(42mg), 60(50mg), 63(3.4g), 67(46m), 72(35mg), 74(2.0g), 79(3mg), 80( 5mg). Fraction 4 was subjected to silica gel column chromatography (petroleum ether: acetone 12:1～3:1), Sephadex LH-20 column chromatography (chloroform-methanol=1:1) to obtain compounds LS-42 (24mg), 52 (28mg ), 54 (14 mg), 66 (13 mg), 68 (49 mg), 70 (38 mg), 76 (34 mg). Fraction 5 was subjected to isogradient elution on silica gel column chromatography (petroleum ether: acetone 4:1) to obtain compounds LS-64 (160 mg) and 65 (62 mg). Fraction 6 was subjected to MCI column chromatography (60% methanol/water solution-pure methanol), and then to silica gel column chromatography (petroleum ether: acetone 4:1) to obtain LS-69 (30mg), 71 (14mg ), 73 (10 mg). Fraction 7 was subjected to Sephadex LH-20 column chromatography (chloroform-methanol=1:1), silica gel column chromatography (dichloromethane: acetone 20:1) and isocratic elution to obtain compounds LS-46 (37 mg), 62 ( 28 mg). Fraction 8 was subjected to MCI column chromatography (60% methanol/water solution-pure methanol), followed by silica gel column chromatography (dichloromethane: acetone 20:1-2:1) gradient elution, and then Sephadex LH-20 Purified by column chromatography (methanol) to obtain compounds LS-44 (19 mg), 45 (29 mg), 48 (19 mg), 49 (29 mg), 55 (25 mg), 58 (30 mg). Fraction 9 was purified by silica gel column chromatography (dichloromethane: acetone 10:1-1:1) gradient elution, and then purified by Sephadex LH-20 column chromatography (methanol) to obtain compound LS-54 (137 mg), 59 (4 mg). Fraction 10 was subjected to MCI column chromatography (50% methanol/water solution-pure methanol), followed by silica gel column chromatography (dichloromethane: acetone 8:1-1:1) gradient elution, and then Sephadex LH-20 Purified by column chromatography (methanol) to obtain compounds LS-50 (13 mg), 56 (73 mg), 57 (85 mg), 77 (12 mg), 78 (9 mg).

The characterization of each compound is as follows:

Fortunin A (LS-41)

white solid. EIMS: m/z 344[M] ⁺ , MF: C ₂₂ H ₃₂ O ₃ , ¹ H NMR (300MHz, CDCl ₃ ) δ7.48(s,1H,H-14),6.82(s,1H,H- 11), 4.86(dd, J=8.6, 8.6Hz, 1H, H-7), 2.89(sept, J=7.0Hz, 1H, H-15), 2.84(m, 2H, H-7), 1.39and 2.15(m,each 1H,H-1),1.41and 1.51(m,each 1H,H-2),1.65and1.75(m,each 1H,H-3),1.39(m,1H,H-5 ), 1.70 and 2.27 (m, each 1H, H-6), 1.20 and 1.22 (sept, J＝6.8Hz, 6H, H-16/17), 0.93, 0.96, 1.25 (s, each 3H, H-18 /19/20),2.32(s,3H,12-CH ₃ CO).

Fortunin B (LS-42)

white solid. EIMS: m/z 402[M] ⁺ , MF: C ₂₄ H ₃₄ O ₅ , ¹ H NMR (300MHz, CDCl ₃ ) δ7.36(s,1H,H-14),6.84(s,1H,H- 11), 5.63 (dd, J=5.3, 11.7Hz, 1H, H-6), 4.69 (brd, J=5.3Hz, 1H, H-7), 2.95 (sept, J=6.8Hz, 1H, H- 15), 1.46 and 2.14 (m, each 1H, H-1), 1.53 (m, 2H, H-2), 1.61 and 1.68 (m, each 1H, H-3), 1.75 (d, J = 11.7 Hz,1H,H-5),1.19and 1.21(sept,J＝6.8Hz,6H,H-16/17),1.02,1.08,1.23(s,each 3H,H-18/19/20),2.32 (s,3H,12-CH ₃ CO),2.18(s,3H,6-CH ₃ CO.

Fortunin C (LS-43)

white solid. EIMS: m/z 362[M] ⁺ , MF: C ₂₂ H ₃₂ O ₄ , ¹ H NMR (600MHz, CDCl ₃ ) δ6.85 (s, 1H, H-14), 4.25 (dd, J＝8.8, 11.1Hz, 1H, H-6), 4.47(d, J＝8.6Hz, 1H, H-7), 3.20(sept, J＝6.8Hz, 1H, H-15), 1.34and 3.07(m, each 1H ,H-1),1.51and 1.68(m,each 1H,H-2),1.28and 1.48(m,each 1H,H-3),1.53(d,J＝11.1Hz,1H,H-5), 1.24(sept,J＝6.8Hz,6H,H-16/17),1.21,1.23,1.43(s,each 3H,H-18/19/20),3.76(s,3H,12-OMe),3.26 (s,3H,7-OMe)

Fortunin D (LS-44)

white solid. EIMS: m/z 362[M] ⁺ , MF: C ₂₂ H ₃₄ O ₄ , ¹ H NMR (600MHz, CDCl ₃ ) δ6.71 (s, 1H, H-14), 4.19 (brd, J = 10.3Hz ,1H,H-6),4.15(d,J=3.8Hz,1H,H-7),3.22(sept,J=7.0Hz,1H,H-15),1.31and 3.01(m,each 1H,H -1),1.49and 1.56(m,each 1H,H-2),1.54and 1.72(m,each 1H,H-3),1.86(d,J＝10.3Hz,1H,H-5),1.25( sept, J＝6.8Hz, 6H, H-16/17), 1.18, 1.20, 1.35(s, each 3H, H-18/19/20), 3.76(s, 3H, 12-OMe), 3.61(s ,3H,7-OMe).

Fortunin E (LS-45)

white solid. EIMS: m/z 332[M] ⁺ , MF: C ₂₀ H ₂₈ O ₄ , ¹ H NMR (300MHz, CDCl ₃ ) δ7.93(s,1H,H-14),6.74(s,1H,H- 11), 4.50(d, J=12.7Hz, 1H, H-6), 3.19(sept, J=7.5Hz, 1H, H-15), 1.36 and 2.17(m, each 1H, H-1), 1.81 (m,2H,H-2),1.47and 1.67(m,each 1H,H-3),2.04(d,J＝12.7Hz,1H,H-5),1.23and 1.25(sept,J＝7.5Hz ,6H,H-16/17),1.16,1.32(s,each 3H,H-18/20),3.26and 3.56(d,J＝11.6Hz,each 1H,H-19).

Fortunin F (LS-46)

Pale yellow solid. EIMS: m/z 330[M] ⁺ , MF: C ₂₀ H ₂₆ O ₄ , ¹ H NMR (300MHz, CDCl ₃ ) δ8.02(s,1H,H-14),6.88(s,1H,H- 11), 3.16(sept, J＝7.0Hz, 1H, H-15), 1.54and 2.24(m, each 1H, H-1), 1.75(m, 2H, H-2), 1.86and 1.96(m, each 1H, H-3), 1.28and 1.30(sept, J＝7.0Hz, 6H, H-16/17), 1.28, 1.50(s, each 3H, H-18/20), 3.35and 4.25(d, J＝11.5Hz, each 1H,H-19).

Fortunin G (LS-47)

white solid. ESIMS: m/z 438.2[M+Na] ⁺ , MF: C ₂₆ H ₃₆ O ₇ , ¹ H NMR (300MHz, CDCl ₃ ) δ7.38(s, 1H, H-14), 6.84(s, 1H, H-11), 5.39(dd, J=5.2, 11.8Hz, 1H, H-6), 4.64(d, J=5.2Hz, 1H, H-7), 2.94(sept, J=6.4Hz, 1H, H-15), 1.52and 1.74(m, each 1H, H-1), 1.68(m, 2H, H-2), 1.42and 1.58(m, each 1H, H-3), 2.13(d, J＝ 11.8Hz, 1H, H-5), 1.20and 1.21(sept, J＝6.4Hz, 6H, H-16/17), 1.00, 1.37(s, each 3H, H-18/20), 3.67and 4.13( d,J＝11.6Hz,each 1H,H-19).

Fortunin H (LS-48)

white solid. EIMS: m/z 316[M] ⁺ , MF: C ₂₀ H ₂₈ O ₃ , ¹ H NMR (600MHz, CDCl ₃ ) δ8.08(s,1H,H-14),7.03(s,1H,H- 11), 4.38(dd, J=7.0, 12.7Hz, 1H, H-6), 5.10(d, J=7.0Hz, 1H, H-7), 3.69(sept, J=7.3Hz, 1H, H- 15), 1.29 and 1.98(m, each 1H, H-1), 1.51(m, 2H, H-2), 1.56 and 1.63(m, each 1H, H-3), 1.68(d, J=12.7 Hz, 1H, H-5), 1.36and 1.37(sept, J＝7.3Hz, 6H, H-16/17), 1.07, 1.11(s, each 3H, H-18/20), 3.47and 3.74(d ,J＝7.4Hz,each 1H,H-19).

Fortunin I (LS-49)

Pale yellow solid. EIMS: m/z 330[M] ⁺ , MF: C ₂₁ H ₃₀ O ₃ , ¹ H NMR (600MHz, CDCl ₃ ) δ7.05(s,1H,H-14),6.57(s,1H,H- 11), 4.17(dd, J=3.8, 12.2Hz, 1H, H-6), 4.46(d, J=3.8Hz, 1H, H-7), 3.15(sept, J=6.3Hz, 1H, H- 15), 1.44 and 2.03(m, each 1H, H-1), 1.75(m, 2H, H-2), 1.30 and 1.82(m, each 1H, H-3), 2.14(d, J=12.2 Hz, 1H, H-5), 1.25and 1.26(sept, J＝6.3Hz, 6H, H-16/17), 1.17, 1.11(s, each 3H, H-18/20), 3.51and 3.77(d ,J＝6.7Hz,each 1H,H-19).

Fortunin J (LS-50)

Pale yellow solid. EIMS: m/z 318[M] ⁺ , MF: C ₂₀ H ₃₀ O ₃ , ¹ H NMR (600MHz, CDCl ₃ ) δ6.76(s,1H,H-14),6.66(s,1H,H- 11), 1.16 and 1.18 (m, each 1H, H-6), 2.51 (dd, J＝4.7, 5.2Hz, 1H, H-7), 3.18 (sept, J＝6.4Hz, 1H, H-15) ,1.26and 2.61(m,each 1H,H-1),4.04(m,1H,H-2),1.47and 1.68(m,each 1H,H-3),1.65(d,J＝4.7,11.0Hz ,1H,H-5),1.17and 1.18(sept,J＝6.4Hz,6H,H-16/17),0.89,1.22(s,each 3H,H-18/20),3.13and 3.44(d, J＝10.8Hz, each 1H,H-19).

Fortunin K (LS-51)

Pale yellow solid. EIMS: m/z 302[M] ⁺ , MF: C ₁₉ H ₂₆ O ₃ , ¹ H NMR (300MHz, CD ₃ OD) δ7.55(s,1H,H-14),6.88(s,1H,H -11), 3.09(sept, J＝6.8Hz, 1H, H-15), 1.52and 2.46(m, each 1H, H-1), 2.00and 2.13(m, each 1H, H-2), 1.68( m,2H,H-3),1.21and 1.23(sept,J＝6.8Hz,6H,H-16/17),1.27,1.30,1.63(s,each 3H,H-18/19/20),9.67 (s,1H,CHO).

Fortunin L (LS-52)

Pale yellow solid. EIMS: m/z 304[M] ⁺ , MF: C ₁₉ H ₂₈ O ₃ , ¹ H NMR (300MHz, CDCl ₃ ) δ7.36(s,1H,H-14),6.84(s,1H,H- 11),1.40and 2.14(m,each 1H,H-1),1.56(m,2H,H-2),1.60(m,2H,H-3),1.25and 1.26(sept,J＝6.8Hz, 6H,H-16/17),0.46,0.88,1.44(s,each 3H,H-18/19/20).

Fortunin M (LS-53)

Pale yellow solid. EIMS: m/z 318[M] ⁺ , MF: C ₂₀ H ₃₀ O ₃ , ¹ H NMR (600MHz, CDCl ₃ ) δ6.72(s,1H,H-14),6.46(s,1H,H- 11), 5.03(d, J=7.4Hz, 1H, H-6), 1.35(d, J=7.4Hz, 1H, H-5), 3.12(sept, J=6.8Hz, 1H, H-15) ,1.26and 2.61(m,each 1H,H-1),4.04(m,1H,H-2),1.47and 1.68(m,each 1H,H-3),1.65(d,J＝4.7,11.0Hz ,1H,H-5),1.22(sept,J=6.4Hz,6H,H-16/17),0.92,1.12,1.23(s,each 3H,H-18/19/20).

Fortunin N (LS-54)

Pale yellow solid. ESIMS: m/z 333.4[M+H] ⁺ , MF: C ₂₀ H ₂₈ O ₄ , ¹ H NMR (400MHz, CD ₃ OD) δ7.81(s,1H,H-14),7.01(s,1H ,H-11),3.11(s,1H,H-5),3.20(sept,J＝6.9Hz,1H,H-15),1.63-2.15(m,6H,H-1/2/3), 1.20(sept,J＝6.4Hz,6H,H-16/17),0.37,0.89,1.43(s,each 3H,H-18/19/20),10.59(s,1H,CHO).

Fortunin O (LS-55)

Pale yellow solid. ESIMS: m/z 319.4[M+H] ⁺ , MF: C ₁₉ H ₂₆ O ₄ , ¹ H NMR (400MHz, CDCl ₃ ) δ7.93(s,1H,H-14),6.77(s,1H, H-11),3.17(sept,J＝6.9Hz,1H,H-15),2.12–1.85(m,1H),1.79–1.58(m,3H),1.57–1.39(m,2H),1.25and 1.26(d,J＝6.9Hz,each 3H,H-16/17),0.68,1.14,1.33(s,each 3H,H-18/19/20).

3-hydroxysugiol (LS-56)

white solid. ESIMS: m/z 317.2[M+H] ⁺ , MF:C ₂₀ H ₂₈ O ₃ , ¹ H NMR (400MHz, CDCl ₃ ) δ7.90(s,1H,H-14),6.68(s,1H, H-11), 3.33(dd, J＝11.0, 4.8Hz, 1H, H-3), 3.16(sept, J＝6.9Hz, 1H, H-15), 2.70–2.66(m, 2H, H-6 ), 1.69and 2.25(m, each 1H, H-1), 1.80-1.94(m, 3H, H-2/5), 1.25and 1.23(d, J＝6.9Hz, each 3H, H-16/17 ),1.21,1.04,0.95(s,each 3H,H-18/19/20).

Cryptojaponol (LS-57)

Pale yellow solid. EIMS: m/z 330[M] ⁺ , MF: C ₂₁ H ₃₀ O ₃ , ¹ H NMR (400MHz, CDCl ₃ ) δ7.60(s, 1H, H-14), 3.18(sept, J＝6.9Hz ,1H,H-15),1.39and 3.23(m,each 1H,H-1),1.57and 1.75(m,each 1H,H-2),1.26-1.48(m,each 1H,H-3), 1.85(m,1H,H-5),2.54and 2.64(m,each 1H,H-6),1.22and 1.24(d,J＝6.9Hz,each 3H,H-16/17),0.92,0.96, 1.32(s, each 3H, H-18/19/20).

Fortunin P (LS-58)

Pale yellow solid. ESIMS: m/z 317.4[M+H] ⁺ , MF: C ₂₀ H ₂₈ O ₃ , ¹ H NMR (400MHz, CDCl ₃ ) δ7.46(s,1H,H-14),6.57(s,1H, H-11), 4.69(d, J=7.7Hz, 1H, H-7), 3.97(dd, J=12.6, 7.7Hz, 1H, H-6), 3.13(sept, J=6.9Hz, 1H) ,2.12-2.05(m,1H,H-5),1.33–2.04(m,6H),1.25and1.27(d,J＝6.9Hz,each 3H,H-16/17),1.19and 1.21(s , each 3H, H-18/20), 3.47 and 3.76 (d, J=7.3Hz, each 1H, H-19), 3.65 (s, 3H, 7-OMe).

Fortunin Q(LS-59)

Pale yellow solid. ESIMS: m/z 333.2[M+H] ⁺ , MF: C ₂₀ H ₂₈ O ₃ , ¹ H NMR (400MHz, CD ₃ OD) δ10.59(s,1H,CHO),7.81(s,1H,H -14),7.01(s,1H,H-11),3.20(sept,J＝6.9Hz,1H,H-15),3.11(s,1H,H-5),1.80-2.15(m,5H) ,1.63(dt,J=14.0,4.2Hz,1H,H-1a),1.20(d,J=6.9Hz,6H,H-16/17),0.37,0.89,1.43(s,each 3H,H- 18/19/20).

6β-hydroxyferruginol (LS-60)

White solid, HREIMS: m/z 302.2247, MF: C ₂₀ H ₃₀ O ₂ . ¹ H-NMR (300MHz, CDCl ₃ ) δ6.82(s,1H),6.66(s,1H),4.65(brs,1H ), 3.10(sept, J=6.8Hz, 1H), 3.08(dd, J=4.5, 17.0Hz, 1H), 2.83(brd, J=17.0Hz, 1H), 2.06(brd, J=12.6Hz, 1H ), 1.37(brs,1H), 1.52(s,3H), 1.25(s,3H), 1.22and 1.21(d,J=6.8Hz, each 3H), 1.02(s,3H).

Iguestol (LS-61)

Pale yellow solid, HREISM m/z 332.2343.MF:C ₂₁ H ₃₂ O ₃ . ¹ H NMR (500MHz, CDCl ₃ ) δ1.13(s,3H,H-18),1.16(s,3H,H-19 ),1.18(d,J=7Hz,3H,H-16),1.20(d,J=7Hz,3H,H-17),1.25(td,J=13,3.3Hz,1H,H-3β), 1.33(s,3H,H-20),1.39(d,1H,J=9Hz,H-5),1.45(dt,J=13.0,4.0Hz,1H,H-3α),1.53(m,2H, H-1R/H-2), 1.70(m, 1H, H-2), 2.74(dd, J＝15.7, 7.6Hz, 1H, H-7), 2.99(dt, J＝13.0, 4.0Hz, 1H ,H-1α),3.14(m,2H,H-7/15),3.72(s,3H-OMe),4.21(1H,brs,J=18Hz,H-6),5.98(s,1H,OH ),6.45(s,1H,H-14).

2-dehydroxy-15-hydroxy iguestol (LS-62)

Pale yellow solid, EISM m/z 332[M] ⁺ .MF:C ₂₁ H ₃₂ O ₃ . ¹ H NMR (300MHz, CDCl ₃ ) δ1.64,1.62,1.37,0.98,0.94(s,each 3H,H -16/17/18/19/20),1.70-1.85(m,3H),1.20-1.54(m,4H),2.48-2.69(m,2H),3.23(m,1H),3.92(s, 3H,12-OMe),5.97(s,1H,11-OH),7.66(s,1H,H-14).

(+)-Sugiol (LS-63)

White powder, EIMS m/z 300(M) ⁺ , MF:C ₂₀ H ₂₈ O ₂ ,[α] _D ²⁵ :+28.3°(c 1.0,CHCl ₃ ), ¹ H NMR(300MHz CDCl ₃ ):δ0. 99,1.00,1.19(each 3H,s,H-18/19/20),1.20(3H,d,J＝7.0Hz,H-16), 1.24(3H,d,J＝7.0Hz,H-17 ), 1.83(dd, J＝4.5, 13.0Hz, H-5), 2.18(dd, J＝13.0, 18.0Hz, H-6β), 2.62(1H, dd, J＝4.5, 1.8Hz, H-6α ), 3.13 (1H, sept, J=7.0Hz, H-15), 6.68 (1H, s, H-11), 7.89 (1H, s, H-14).

11-hydroxysugiol (LS-64)

Pale yellow powder, EIMS m/z 316(M) ⁺ ,MF:C ₂₀ H ₂₈ O ₃ , ¹ H NMR(300MHz CDCl ₃ ):δ0.92,0.97,1.40(each 3H,s,H-18/19 /20),1.22and 1.24(each 3H,d,J＝7.0Hz,H-16/17),1.87(dd,J＝3.2,14.2Hz,H-5),2.53(dd,J＝14.2,17.4 Hz,H-6β),2.62(1H,dd,J＝14.2,17.4Hz,H-6α),1.28and 1.50(each 1H,m,H-3),1.78and 1.61(each 1H,m,H- 2), 1.51 and 3.11 (each 1H, m, H-1), 3.13 (1H, sept, J = 7.0Hz, H-15), 5.81 and 6.13 (2H, brs, 11/12-OH), 7.63 ( 1H, s, H-14).

Hypagenin B (LS-65)

Pale yellow powder, EIMS m/z 316(M) ⁺ ,MF:C ₂₀ H ₂₈ O ₃ , ¹ H NMR(300MHz CDCl ₃ ):δ7.58(s,lH,H-14),6.47(s,1H ,H-11),3.0(m,1H,H-6b)2.8(br d,J=12Hz,1H,H-1a),2.4(dd,J=4,12Hz,1H,H-6a),1.4 -2.4(m,6H),1.40,1.35,1.12,1.10,1.08(s,each 3H,H-16/17/18/19/20).

5,6-dehydrosugiol (LS-66)

Pale yellow powder, HREIMS: 298.1933[M+H] ⁺ , MF: C ₂₀ H ₂₆ O ₂ , ¹ H NMR (400MHz, CDCl ₃ ): δ＝8.01(s,1H),7.15(s,1H),6.84 (s,1H),5.24(s,1H),3.17(sept,J＝6.8Hz,1H),2.33-2.27(m,1H),1.97-1.87(m,2H),1.81-1.70(m,2H ),1.51,1.43,1.43(s,each 3H),1.31and 1.28(d,J＝6.8Hz,each 3H),1.25-1.23(m,1H).

6-hydroxy-5,6-dehydrosugiol (LS-67)

Pale yellow powder, EIMS: 314[M] ⁺ , MF: C ₂₀ H ₂₆ O ₃ , ¹ H NMR (400MHz, DMSO-d ₆ ) δ1.16(d, J=6.8Hz, 3H), 1.18(d, J＝6.8Hz, 3H), 1.19(s, 3H), 1.29(s, 3H), 1.42(s, 3H), 1.31–2.30(m, 6H), 3.16(sept., J＝6.8Hz, 1H) ,6.23(s,1H),6.94(s,1H),7.71(s,1H),10.20(s,1H).

14-Deoxycoleon U(LS-68)

Light yellow powder, EIMS:330[M] ⁺ ,MF:C ₂₀ H ₂₆ O ₄ ,1H NMR(500MHz,CDCl ₃ )δ1.27and 1.30(each 3H,d,J＝6.8Hz,H-16/17) ,1.41(1H,m,H-3α),1.43and 1.44(each 3H,s,H-18/19),1.61and 1.86(each 1H,m,H-2),1.71and 2.93(each 1H,m ,H-1),2.01(1H,td,J=13,5Hz,H-3β),3.04(1H,sept,J=6.8Hz,H-15),7.09(1H,s,6-OH), 7.70(1H,s,H-14).

12-O-Acetylhinokiol (LS-69)

Light yellow powder, EIMS:344[M] ⁺ ,MF:C ₂₂ H ₃₂ O ₃ , ¹ H NMR(500MHz,CDCl ₃ )δ1.11and 1.36(3H,m),1.58-1.93(5H,m),1.18 ,1.05and 0.89(each 3H,s,H-18/19/20),2.20(1H,m),2.30(3H,s,OMe),2.90(1H,sept,J＝6.8Hz,H-15) ,3.28(1H,m,H-3),6.81(1H,s,H-11),6.95(1H,s,H-14).

3β-Acetoxyabieta-8,11,13-trien-12-ol (LS-70)

Pale yellow solid, EIMS m/z 344[M ⁺ ], MF:C ₂₂ H ₃₂ O ₃ , ¹ H NMR(CDCl ₃ ,400MHz)δ6.81(1H,s,H-14),6.58(1H,s ,H-11),5.03(1H,brs,OH),4.51(1H,dd,J=11.1,4.9Hz,H-3),3.09(1H,sept,J=6.8Hz,H-15),2.83 (1H,ddd,J=12.6,4.0,1.5Hz,Hα-7),2.75(1H,ddd,J=12.6,11.3,3.5Hz,Hβ-7),2.17(1H,ddd,J=13.2,3.3 ,3.3Hz,Hα-1),2.05(3H,s,OAc),1.60(1H,ddd,J＝13.2,11.0,3.0Hz,Hβ-1),1.36(1H,dd,J＝12.0,2.1Hz ,H-5),1.21,1.20(3H each,d,J＝6.8Hz,H-16/17),1.18(3H,s,H-20),0.94(3H,s,H-19),0.92 (3H,s,H-18).

Salviviridinol (LS-71)

Pale white solid, EIMS m/z 348[M ⁺ ], MF:C ₂₁ H ₃₂ O ₄ , ¹ H NMR(CDCl ₃ ,400MHz)δ6.99(1H,s,H-14),4.49(1H,d ,J=9.5Hz,H-7),3.98(1H,d,J=9.5,13.0Hz,H-6),3.20(1H,sept,J=6.8Hz,H-15),3.08and 1.70(each 1H,m,H-1),1.40and 1.60(each 1H,m,H-2),1.60and 1.80(each 1H,m,H-3),1.52(1H,d,J＝13.0Hz,H- 5),1.17(6H,d,J＝6.8Hz,H-16/17),1.10,1.24,1.04(each 3H,s,H-18/19/20),3.80(3H,s,OMe).

Sugikurojin D (LS-72)

White solid, EIMS m/z 390[M ⁺ ], MF:C ₂₃ H ₃₄ O ₅ , ¹ H NMR(CDCl ₃ ,400MHz)δ7.03(1H,s,H-14),4.6149(1H,d, J＝8.5Hz, H-7), 5.42(1H, d, J＝8.5, 12.0Hz, H-6), 3.19(1H, sept, J＝7.0Hz, H-15), 3.15and 1.32(each 1H ,m,H-1),1.51and 1.68(each 1H,m,H-2),1.28and 1.45(each 1H,m,H-3),1.73(1H,d,J＝12.0Hz,H-5 ),1.24and 1.21(each 3H,d,J＝7.0Hz,H-16/17),1.15,0.99,1.49(each 3H,s,H-18/19/20),3.76(3H,s,OMe ), 2.19(3H,s,Ac).

18-Hydroxyferruginol (LS-73)

White solid, EIMS m/z 302[M ⁺ ], MF:C ₂₀ H ₃₀ O ₂ , ¹ H NMR(600MHz,CDCl ₃ )δ0.86(3H,s,H-19),1.18(3H,s, H-20),1.21(3H,d,H-16),1.22(3H,d,H-17),1.36(1H,m,H-1α),1.42(1H,m,H-3),1.59 (1H,m,H-5),1.64(1H,m,H-6),1.71(1H,m,H-2),2.16(1H,m,H-1β),2.80(2H,m,H -7),3.09(1H,m,H-15),3.21(1H,d,J=10.9Hz,H-18α),3.45(1H,d,J=11.0Hz,H-18β),6.61(1H ,s,H-11),6.81(1H,s,H-14).

12-hydroxy-6,7-seco-abieta-8,11,13-triene-6,7-dial (LS-74)

White solid, HREIMS m/z 316.2038, MF: C ₂₀ H ₂₈ O ₃ , ¹ H NMR (400MHz, CDCl ₃ ) δ10.47(s, 1H), 9.86(d, J=3.8Hz, 1H), 7.84( s,1H),6.96(s,1H),6.34(br s,1H),3.18(sept,J＝6.9Hz,1H),3.13(d,J＝3.8Hz,1H),2.28–2.34(m, 1H), 1.50–1.83(m,5H), 1.50(s,3H), 1.27and 1.26(d, J＝6.9Hz, each 3H), 1.01and 0.70(s, each 3H).

Cupresol (LS-75)

White solid, EIMS m/z 318[M] ⁺ , MF:C ₁₉ H ₂₆ O ₄ , ¹ H NMR (300MHz, CDCl ₃ )δ7.96(s,1H,H-14),6.76(s,1H, H-11),5.82-5.99(br s,2H,OH),3.18(sept,J=6.9Hz,1H,H-15),3.13(m,1H,H-1a),1.39–2.08(m, 5H), 1.27and 1.26(d, J＝6.9Hz, each 3H, H-16/17), 1.33, 1.14and 0.70(s, each 3H).

7β-hydroxydeoxy-cryptojaponol (LS-76)

Pale white solid, EIMS m/z 332[M] ⁺ , MF:C ₂₁ H ₃₂ O ₃ , ¹ H NMR (300MHz, CDCl ₃ ) δ6.63(s,1H,H-14), 4.41(1H,m ,H-7),1.55and 5.72(br s,2H,OH),2.96(sept,J=6.9Hz,1H,H-15),3.13(m,1H,H-1a),1.31–2.07(m ,6H),1.27and 1.15(d,J＝6.9Hz,each 3H,H-16/17),0.93,0.93and 1.35(s,each 3H,H-18/19/20),3.58(s,3H ,OMe).

Fortunin R (LS-77)

White color solid. ESIMS: m/z 349.4[M+H]+, MF: C ₂₀ H ₂₈ O ₅ , 1H NMR (400MHz, CD ₃ OD) δ8.06(s,1H,H-14),6.36(S,1H, H-11), 3.69(dd, J=10.3, 5.7Hz, 1H, H-16a), 3.54(dd, J=11.7, 4.9Hz, 1H, H-1), 3.48(dd, J=10.2, 7.4 Hz, 1H, H-16b), 3.14(m, 1H, H-15), 2.66(m, 2H, H-7), 2.21(dt, J=13.2, 3.5Hz, 1H, H-3a), 2.07 (m,1H,H-2α),2.05(m,2H,H-6),1.70(dt,1H,J=13.6,3.6Hz,H-3b),1.25(s,3H,H-19), 1.24(d, J=6.8Hz, 3H, H-17), 1.18(m, 1H, H-3b), 1.13(s, 3H, H-20).

Fortunin S (LS-78)

white solid. ESIMS: m/z 349.4[M+H]+, MF: C ₂₀ H ₂₈ O ₅ , 1H NMR (400MHz, CD ₃ OD) δ6.71(s,1H,H-14),6.65(s,1H, H-11), 4.05 (brs, 1H, H-3), 3.68 (dd, J＝10.5, 5.8Hz, 1H, H-16a), 3.49 (dd, J＝10.4, 7.4Hz, 1H, H-16b ),3.16(m,1H,H-15),2.73(m,2H,H-7),2.30(m,1H,H-2a),2.00(m,2H,H-2α),1.90(m, 1H,H-5),1.89(m,1H,H-1a),1.74(m,1H,H-1b),1.70(m,1H,H-2b),1.31(s,3H,H-19) ,1.21(d,J=6.8Hz,3H,H-17),1.09(s,3H,H-20).

Fortunin T (LS-79)

white solid. EIMS: m/z 330[M]+, MF: C ₂₁ H ₃₀ O ₃ , 1H NMR (400MHz, CDCl3) δ9.81(s, 1H, CHO-), 7.14(s, 1H, H-14), 6.48(s,1H,H-11),5.30(s,1H,OH-Ar),4.70(s,1H,H-7),3.52(s,3H,7-OMe),3.19(sept,J= 6.8Hz, 1H, H-15), 1.23(d, J＝6.8Hz, 6H, H-16/17), 1.43-1.96(m, 6H, H-1/2/3), 1.25, 1.11, 0.53 (s, each 3H, H-20/19/18).

Fortunin U (LS-80)

white solid. EIMS: m/z 304[M]+, MF: C19H28O3, 1H NMR (400MHz, Acetone-d6) δ7.94(s, 1H, OH-Ar), 7.13(s, 1H, H-14), 6.54( s,1H,H-11),5.12(d,J=7.2Hz,1H,H-7),4.19(d,J=7.2Hz,1H,7-OH),3.24(sept,J=6.8Hz, 1H,H-15),3.16(s,1H,5-OH),1.65–1.20(m,6H,H-1/2/3),1.16and 1.18(d,J＝6.8Hz,3H,H- 16/17), 1.36, 1.10, 1.01(s, each 3H, H-20/19/18).

1.2. Separation of abietane-type diterpenoids from the bark of Japanese fir

Grind the bark (27Kg) of Pondus fir collected from Yiyang, Hunan, and soak it in industrial acetone at room temperature for three times, each time for 7 days. Dichloromethane and ethyl acetate were extracted three times each, and concentrated under reduced pressure to obtain 118 g of petroleum ether, 180 g of dichloromethane, and 1.2 Kg of ethyl acetate. The dichloromethane part is passed through normal phase silica gel (100-200 mesh, 200-300 mesh, silica gel H, etc.), elution conditions: petroleum ether/acetone, chloroform/acetone, chloroform/methanol), Sephadex LH-20 (chloroform/methanol =1:1, methanol), MCI resin (methanol/water, ethanol/water), preparative thin-layer and preparative high-performance liquid phase column chromatography for separation, a total of 22 compounds were separated and identified, 13 of which were new compounds, including 3 abietane-type diterpenes Taxodascen A (7,18mg), Taxodascen B (10,13mg), Taxodascen C (11,31mg) and 1 abietane-type Taxodascen D (12,25mg) with B ring cleavage, 3 pcs A ring-cleaved abietane-type diterpenes Nortaxodascen A (13,20mg), Nortaxodascen B (15,35mg), Nortaxodascen C (16,51mg), dimer of 6 abietane diterpenes Bitaxodascen A (17,50mg) , Bitaxodascen B (18, 12mg), Bitaxodascen C (19, 8mg), Bitaxodascen D (20, 27mg), Bitaxodascen E (21, 85mg), Bitaxodascen F (22, 70mg). 9 known compounds Ferruginol (1,85mg), Abieta-8,11,13-trien-12,19-diol (2,80mg), 6,7-dehydroferruginol (3,126mg), 19-hydroxy-6,7 -Dehydroferruginol (4,12mg), sugiol (5,1.7g), 6α-hydroxy-7-oxoferruginol (6,65mg), Fortunin E (8,14mg), 6,12-dihydroxyabieta-5,8,11,13 -tetraen-7-one(9,141mg), 20(10->5)abeo-4,5-secoabieta-5(10),6,8,11,13-pentaene-3,4,12-triol(14 ,34mg).

The characteristics of each compound are as follows:

Ferruginol(1)

Pale yellow gelatinous solid. ESIMS: m/z 287.0[M+H] ⁺ , MF: C ₂₀ H ₃₀ O, ¹ H NMR (400MHz, CDCl ₃ ) δ6.83(s,1H,H-14),6.63(s,1H,H -11),4.55(s,1H,12-OH),3.11(sept,J＝7.0Hz,1H,H-15),2.84(m,2H,H-7),1.90-1.80(m,1H) ,1.78-1.70(m,1H),1.69-1.63(m,1H),1.64-1.55(m,2H),1.50-1.43(m,1H),1.40(dd,J＝13.0,3.9Hz,1H) ,1.35-1.28(m,1H),1.23(t,J＝6.8Hz,6H,H-16/17),1.17(s,3H,H-20),0.94(s,3H,H-18), 0.92(s,3H,H-19).

Abieta-8,11,13-trien-12,19-diol (2)

Yellow amorphous powder. ESIMS: m/z 300.9[MH] ^- , MF: C ₂₀ H ₃₀ O ₂ , ¹ H NMR (400MHz, CDCl ₃ ) δ6.82(s,1H),6.63(s,1H),3.86(d,J =11.0Hz,1H),3.55(d,J=11.0Hz,1H),3.11(sept,J=6.9Hz,1H),2.90-2.69(m,2H),2.04-1.82(m,2H),1.75 -1.55(m,3H),1.52-1.35(m,2H),1.23(t,J＝6.6Hz,6H),1.16(s,3H),1.05(s,3H).

6,7-dehydroferruginol (3)

Pale yellow gelatinous solid. ESIMS: m/z 283.0[MH] ^- , MF: C20H28O, ¹ H NMR (400MHz, CDCl ₃ ) δ6.89 (d, J = 8.0Hz, 1H), 6.58 (s, 1H), 6.49 (dd, J =9.6,3.1Hz,1H),5.87(dd,J=9.6,2.7Hz,1H),4.84(s,1H),3.12(sept,J=6.9Hz,1H),2.07(dd,J=6.0, 3.1Hz, 2H), 1.66-1.57(m, 3H), 1.54-1.47(m, 2H), 1.27(dd, J=6.9Hz, 4H), 1.22(dd, J=6.9Hz, 3H), 1.04( s,3H),1.01(s,3H),0.96(s,3H).

19-hydroxy-6,7-dehydroferruginol (4)

Pale yellow powdery solid. ESIMS: m/z 298.9301.0[M+H] ⁺ , MF: C ₂₀ H ₂₈ O ₂ , ¹ H NMR (300MHz, CDCl ₃ ) δ6.88(s,1H),6.59(s,1H),6.48 (dd, J=9.6,3.0Hz,1H),5.96(dd,J=9.6,2.3Hz,1H),3.85(d,J=11.2Hz,1H),3.73(d,J=11.3Hz,1H) ,3.15(sept,J=6.9Hz,1H),2.24(s,1H),2.10(d,J=11.6Hz,1H),1.88(d,J=13.9Hz,2H),1.25(d,J= 6.9Hz, 4H), 1.22(d, J=6.9Hz, 3H), 1.06(s, 4H), 0.98(s, 3H).

sugiol (5)

Pale yellow granular crystal, slightly soluble in chloroform, acetone, methanol and other organic solvents. ESIMS: m/z 301.3[M+H] ⁺ , MF: C ₂₀ H ₂₈ O ₂ , ¹ H NMR (300MHz, DMSO-d6) δ10.25(s, 1H), 7.66(s, 1H), 6.80( s,1H),3.14(sept,J=6.9Hz,1H),2.23–2.05(m,1H),1.75(dd,J=13.0,4.1Hz,2H),1.63(t,J=14.4Hz,1H ), 1.43(t, J=15.6Hz, 2H), 1.34–1.21(m, 1H), 1.16(overlap, 9H), 0.95(s, 3H), 0.89(s, 3H).

6α-hydroxy-7-oxoferruginol (6)

Pale yellow colloidal solid, slightly soluble in chloroform, acetone, methanol and other organic solvents. ESIMS: 316.94[M+H] ⁺ , MF: C ₂₀ H ₂₈ O ₃ , ¹ H NMR (400MHz, CDCl ₃ ) δ7.96(s,1H),6.72(s,1H),5.71(s,1H) ,4.65(dd,J=12.8,1.6Hz,1H),3.95(d,J=1.8Hz,1H),3.17(sept,J=6.9Hz,1H),2.20(t,J=5.3Hz,1H) ,1.85(d,J=12.8Hz,1H),1.77(tt,J=13.4,3.1Hz,1H),1.64(dt,J=14.0,3.7Hz,2H),1.56–1.48(m,2H), 1.38(s,3H),1.28(t,J=6.9Hz,6H),1.25(s,3H),1.22(s,3H).

Taxodascen A(7)

Pale yellow gelatinous solid. ESIMS: m/z 315.3[M+H] ⁺ ,MF:C ₂₀ H ₂₆ O ₃ , ¹ H NMR (300MHz,acetone-d ₆ )δ8.00(s,1H),6.91(s,1H),6.46 (s,1H),3.90(d,J=11.5Hz,1H),3.57(d,J=11.5Hz,1H),3.22(sept,J=6.9Hz,1H),2.31(m,1H),1.90 (m,1H),1.83(m,1H),1.68(m,1H),1.62(m,1H),1.51(s,3H),1.36(m,1H),1.42(s,3H),1.24( s,3H),1.26(dd,J=6.9Hz,6H).

Fortunin E(8)

Yellow granular solid. ESIMS: m/z 331.0[M+H]+, MF: C ₂₀ H ₂₆ O ₄ , ¹ H NMR (300MHz,acetone-d ₆ )δ7.92(s,1H),7.09(s,1H),4.15 (d,J=10.5Hz,1H),3.59(d,J=10.5Hz,1H),3.31(sept,J=6.9Hz,1H),2.48–2.24(m,2H),1.51(s,3H) ,1.35(s,3H),1.26(dd,J=6.9Hz,6H).

6,12-dihydroxyabieta-5,8,11,13-tetraen-7-one(9)

Pale yellow powdery solid. ESIMS: m/z 314.9[M+H] ⁺ , MF: C ₂₀ H ₂₆ O ₃ , ¹ H NMR (400MHz, CDCl ₃ ) δ8.01(s,1H,H-14),7.15(s,1H) ,6.86(s,1H,H-11),3.18(sept,J=7.0Hz,1H H-15,),2.27(dd,J=13.9,5.9Hz,1H,H-1β),2.00–1.82( m, 2H), 1.80–1.67 (m, 2H), 1.49 (s, 3H, H-20), 1.43 (s, 6H, H-18 and H-19), 1.28 (d, J=6.9Hz, 3H ,H-16),1.26(d,J=6.9Hz,3H,H-17).

Taxodascen B(10)

Pale yellow solid powder. MF:C20H28O4,ESIMS:m/z 331.0[MH] ^- ,[α] ²² _D 52.3(c 0.13,MeOH). ¹ H NMR(400MHz,CDCl ₃ )δ6.79(s,1H,H-11), 6.77(s,1H,H-14),4.20(t,J=11.4,4.4Hz,1H,H-2),3.20(sept,J=7.0Hz,1H,H-15),2.79(dd,J =16.3,4.3Hz,1H,H-7a),2.68(m,1H,H-7b),2.49(m,1H,H-1a),2.47(m,1H,H-3a),2.17(m, 1H, H-6a), 1.97(m, 1H, H-6b), 1.24(m, 1H, H-1b), 1.33(s, 3H, H-18), 1.16(d, 3H, J=6.8Hz ,H-16),1.18(d,3H,J＝6.8Hz,H-17),1.08(s,3H,H-20).

Taxodascen C(11)

Pale yellow gelatinous solid. MF: C ₂₀ H ₂₆ O ₄ , ESIMS: m/z 328.9[MH] ^- , ¹ H NMR (400MHz, CDCl ₃ ) δ7.52(s,1H,H-11),6.60(s,1H,H- 14), 5.16(d, J=3.2Hz, 1H, H-7), 4.85(dd, J=5.8, 3.2Hz, 1H, H-6), 3.19(sept, J=7.0Hz, 1H, H- 15), 2.26(dd, J=14.5, 6.0Hz, 1H, H-3a), 1.46(dd, J=13.3, 8.8Hz, 1H, H-3b), 1.96(m, 1H, H-1a), 1.61(m,1H,H-1b),1.81(m,1H,H-2a),1.68(m,1H,H-2b),1.81(d,J=5.8Hz,1H,H-5),1.28 (s,3H,H-18),1.26(d,3H,J=6.8Hz,H-16),1.23(d,3H,J=6.8Hz,H-17),1.02(s,3H,H- 20).

Taxodascen D(12)

Pale yellow colloidal solid. MF: C ₂₀ H ₂₈ O ₄ , ESIMS: m/z 330.9[MH] ^- . ¹ H NMR (400MHz, CDCl ₃ ) δ7.88(s, 1H, H-11), 7.15 (s,1H,H-14),10.70(s,1H,H-7),3.28(sept,J=7.0Hz,1H,H-15),3.20(s,1H,H-5),2.01( m,1H,H-3a),1.16(m,1H,H-3b),2.11(m,1H,H-1a),1.98(m,1H,H-1b),1.84(m,1H,H-1b) 2a), 1.62(m, 1H, H-2b), 0.93(s, 3H, H-18), 1.24(d, 3H, J=6.9Hz, H-16), 1.23(d, 3H, J=6.9 Hz,H-17),0.43(s,3H,H-19),1.02(s,3H,H-20).

Nortaxodascen A(13)

Pale red gelatinous solid. HRESIMS m/z 271.1304[MH]-, MF: C ₁₇ H ₂₀ O ₃ . ¹ H NMR (400MHz, CDCl ₃ ) δ7.59(s,1H,H-11),7.29(s,1H,H-14 ),7.54(d,J=8.3Hz,1H,H-7),7.11(d,J=8.3Hz,1H,H-6),3.37(sept,J=7.0Hz,1H,H-15), 3.20(s,1H,H-5),3.34(m,1H,H-1),2.66(m,1H,H-2),1.34(d,6H,J＝6.9Hz,H-16,17) ,2.46(s,3H,H-20).

20(10->5)abeo-4,5-secoabieta-5(10),6,8,11,13-pentaene-3,4,12-triol(14)

Orange-red powdery solid. ESIMS: m/z 314.9[MH]-, MF: C ₂₀ H ₂₈ O ₃ , ¹ H NMR (300MHz, CDCl ₃ ) δ7.58(s, 1H), 7.52(d, J=8.3Hz, 1H), 7.43(s,1H),7.09(dd,J＝8.2,2.7Hz,1H),3.69–3.61(m,1H),3.46–3.26(m,2H),3.03–2.89(m,1H),2.44( s,3H),1.74(m,2H),1.34(d,J=6.9Hz,6H),1.24(s,3H),1.17(s,3H).

Nortaxodascen B(15)

Orange-red powdery solid. MF: C ₂₀ H ₂₈ O ₃ . ESIMS: m/z 314.9[MH]-, ¹ H NMR (400MHz, CDCl ₃ ) δ7.56(s, 1H, H-11), 7.50 (d,J=8.3Hz,1H,H-7),7.30(s,1H,H-14),7.08(d,J=8.3,1H,H-6),3.38(sept,J=6.9Hz, 1H, H-15), 3.52(d, J=11.4Hz, H-18a), 3.44(d, J=11.4Hz, H-18b), 2.91(t, J=7.9Hz, 2H, H-1) ,167(m,2H,H-2),1.76(m,2H,H-3),1.32(d,J=6.8Hz,6H,H-16,17),1.11(s,3H,H-19 ),2.41(s,3H,H-20).

Nortaxodascen C(16)

Orange-red gelatinous solid. MF: C ₂₀ H ₂₆ O ₃ .ESIMS: m/z 312.9[MH] ^- , ¹ H NMR (400MHz, CDCl ₃ ) δ7.56(s, 1H, H-11), 7.50(d, J＝8.3Hz ,1H,H-7),7.24(s,1H,H-14),7.10(d,J=8.3,1H,H-6),3.35(sept,J=6.9Hz,1H,H-15), 3.02(m,1H,H-1a),2.90(m,1H,H-1b),1.67(m,2H,H-2b),1.90(m,1H,H-3a),1.68(m,1H, H-3b), 2.45(m, 1H, H-4), 1.33(d, J=6.8Hz, 6H, H-16, 17), 1.22(d, J=6.8Hz, 3H, H-18), 2.42(s,3H,H-20).

Bitaxodascen A(17)

Pale yellow granular crystals. [α] ²² _D 90.0(c 0.08,MeOH),ESIMS:m/z 629[MH] ^- , ¹ H NMR(600MHz,CDCl ₃ )δ1.55and 1.28(m,each 1H,H-1),1.61( m,2H,H-2),1.88and 1.09(m,each1H,H-3),1.71(d,J＝8.6Hz,1H,H-5),5.35(d,J＝8.6Hz,1H,H -6),3.97(s,H-7),6.87(s,1H,H-11),6.96(s,1H,H-14),3.17(sept,J＝6.9Hz,H-15),1.16 and 1.20(d,J＝6.9Hz,each 3H,H-16,17),1.02and 1.40(s,each 3H,H-18,20),3.89(m,2H,H-19),3.14(s ,3H,7-OMe); 2.27and 1.69(m,each 1H,H-1"),1.69and 1.80(m,each 1H,H-2"),1.03and 2.02(m,each 1H,H-3 "), 2.21(dd, J＝3.0, 3.0Hz, 1H, H-5"), 6.05(dd, J＝9.6, 3.0Hz, 1H, H-6"), 6.51(dd, J＝9.6, 3.0 Hz,1H,H-7"),7.11(s,1H,H-11"),6.96(s,1H,H-14"),3.27(sept,J＝6.9Hz,1H,H-15") ,1.03and 1.08(d,J＝6.9Hz,each 3H,H-16",17"),1.06(s,6H,H-18",20"),3.63and 3.84(d,J＝10.8Hz, H-19"a,19"b).

Bitaxodascen B(18)

Pale yellow powder crystal. MF:C ₄₀ H ₅₆ O ₅ ,ESIMS:m/z 615.43[MH] ^- , ¹ H NMR(600MHz,CDCl ₃ )δ1.59and 2.16(m,each 1H,H-1),1.59and 1.74(m, each 1H,H-2),1.05and 1.97(m,each 1H,H-3),1.77(d,J＝9.1Hz,1H,H-5),5.25(d,J＝9.1Hz,1H,H -6),4.59(s,H-7),6.81(s,1H,H-11),6.97(s,1H,H-14),3.23(sept,J＝6.8Hz,H-15),1.16 and 1.14(d, J＝6.8Hz, each 3H, H-16,17), 1.04and 1.47(s, each 3H, H-18, 20), 3.82and 3.98(d, J＝10.7Hz, each 1H, H-19);2.34and 1.66(m,each 1H,H-1"),1.64and 1.80(m,each 1H,H-2"),1.02and 1.99(m,each 1H,H-3"), 2.20(dd,J＝3.0,3.0Hz,1H,H-5"),6.03(dd,J＝9.6,3.0Hz,1H,H-6"),6.50(dd,J＝9.6,3.0Hz,1H ,H-7"),7.26(s,1H,H-11"),6.94(s,1H,H-14"),3.23(sept,J＝6.9Hz,1H,H-15"),1.06and 1.10(d,J＝6.9Hz, each 3H,H-16",17"),1.03and 1.05(s,each 3H,H-18",20"),3.61and 3.85(d,J＝10.9Hz, H-19"a,19"b).

Bitaxodascen C(19)

Pale yellow gelatinous solid. ₍ ^{_ _} _{_} m,each 1H,H-2),1.19and1.85(m,each 1H,H-3),2.36(d,J＝11.3Hz,1H,H-5),6.03(d,J＝11.3Hz, 1H,H-6),4.59(s,H-7),6.97(s,1H,H-11),7.69(s,1H,H-14),3.27(sept,J＝6.9Hz,H-15 ),1.20and 1.21(d,J＝6.8Hz,each 3H,H-16,17),1.14and 1.42(s,each 3H,H-18,20),3.81and 3.94(m,each 1H,H- 19);2.54and 1.85(m,each 1H,H-1"),1.62and 1.70(m,each 1H,H-2"),1.38and 1.89(m,each 1H,H-3"),6.31( s,1H,H-6"),7.48(s,1H,H-11"),7.92(s,1H,H-14"),3.39(sept,J＝6.9Hz,1H,H-15") ,1.17and 1.22(d,J＝6.9Hz,each 3H,H-16",17"),1.24and 1.51(s,each 3H,H-18",20"),3.63and 3.83(m,each 1H ,H-19"a,19"b).

Bitaxodascen D(20)

Yellow powdery solid,[α] ²² _D 128.6(c 0.08,MeOH).MF:C ₄₀ H ₅₂ O ₄ .ESIMS:m/z 595.1[MH] ^- , ¹ H NMR(600MHz,CDCl ₃ )δ1.66and 2.17(m, each 1H, H-1), 1.72(m, 2H, H-2), 1.10 and 1.94(m, each 1H, H-3), 1.77(d, J＝7.6Hz, 1H, H- 5), 5.20(dd, J=7.6, 7.3Hz, 1H, H-6), 4.87(d, J=7.4Hz, 1H, H-7), 6.66(s, 1H, H-11), 6.69( s, 1H, H-14), 2.92(sept, J＝6.9Hz, H-15), 1.14and1.18(d, J＝6.8Hz, each 3H, H-16, 17), 1.21and 1.28(s ,each 3H,H-18,20),3.87and 4.01(d,J＝11.0Hz,each 1H,H-19);6.32(s,1H,H-1"),2.08and 2.24(m,each 1H ,H-2"),1.43and1.66(m,each 1H,H-3"),6.06(s,1H,H-6"),6.31(s,1H,H-7"),6.73(s ,1H,H-14"),2.97(sept,J=6.9Hz,1H,H-15"),1.17(d,J=6.9Hz,6H,H-16",17"),1.04and 1.40( s, each 3H, H-18", 20"), 3.63 and 3.83 (d, J＝11.0Hz, each 1H, H-19"a, 19"b).

Bitaxodascen E(21)

Light yellow powdery solid, [α] ²² _D 245.7 (c 0.07, MeOH), MF: C ₄₀ H ₅₄ O ₅ .ESIMS: m/z 613.2[MH] ^- . ¹ H NMR (600MHz, CDCl ₃ ) δ1. 48and 2.14(m, each 1H, H-1), 1.66and 1.71(m, each1H, H-2), 1.14and 1.82(m, each 1H, H-3), 1.89(d, J＝7.4Hz, 1H ,H-5),5.53(dd,J=7.4,7.8Hz,1H,H-6),4.72(d,J=7.4Hz,1H,H-7),6.64(s,1H,H-11) ,6.51(s,1H,H-14),2.89(sept,J=6.8Hz,H-15),0.70and 0.85(d,J=6.8Hz,each 3H,H-16,17),1.25and 1.34 (s, each 3H, H-18, 20), 3.87 and 4.01 (d, J＝11.0Hz, each 1H, H-19); 2.84 and 2.99 (m, each 1H, H-1"), 1.47 and 1. 66(m,each 1H,H-2"),1.47and 2.15(m,each 1H,H-3"),7.13(d,J＝8.4Hz,1H,H-6"),7.57(d,J =8.4Hz,1H,H-7"),7.48(s,1H,H-14"),3.11(sept,J=6.9Hz,1H,H-15"),1.26and 1.27(d,J=6.9 Hz, each 3H, H-16", 17"), 1.03, 2.47 and 3.33(s, each 3H, H-18", 19", 20").

Bitaxodascen F(22)

Yellow amorphous powder. MF:C ₄₀ H ₅₂ O ₄ ,[α] ²² _D 213.3(c 0.07,MeOH),ESIMS:m/z 594.92[MH] ^- . ¹ H NMR(600MHz,CDCl ₃ )δ1.70and 2.21(m,each 1H,H-1),1.71and 1.79(m,each1H,H-2),1.72and 2.19(m,each 1H,H-3),1.93(d,J＝7.8Hz,1H,H-5), 5.48(t, J=7.8Hz, 1H, H-6), 4.81(d, J=7.8Hz, 1H, H-7), 6.68(s, 1H, H-11), 6.57(s, 1H, H -14), 2.91(sept, J＝6.8Hz, H-15), 0.74and 0.89(d, J＝6.8Hz, each 3H, H-16, 17), 1.31and 1.32(s, each 3H, H- 18,20),4.00and 4.04(d,J＝10.8Hz,each 1H,H-19);2.97and 3.09(m,each 1H,H-1"),2.27(m,2H,H-2") ,5.39(t,J＝6.6Hz,1H,H-3"),7.18(d,J＝8.3Hz,1H,H-6"),7.62(d,J＝8.3Hz,1H,H-7"),7.52(s,1H,H-14"),3.16(sept,J＝6.9Hz,1H,H-15"),1.29and 1.30(d,J＝6.9Hz, each3H,H-16",17 "),1.60,2.52 and 3.93(s,each 3H,H-18",19",20").

1.3. Separation of abietane-type diterpenoids in the fruit of Pondus chinensis

Pulverize the Chishan fruit (25Kg) collected from She County, Anhui, soak it three times with 95% industrial ethanol at room temperature, each time for 7 days, combine the extracts, concentrate under reduced pressure to remove ethanol, add 12L to suspend, and use petroleum ether, Dichloromethane and ethyl acetate were extracted three times each, and concentrated under reduced pressure to obtain 120 g of petroleum ether, 200 g of dichloromethane, and 47 g of ethyl acetate. After the dichloromethane part is eluted with 30%, 60%, 80%, 95% ethanol solution and acetone of MCI resin, the obtained fractions are sequentially passed through normal phase silica gel (100-200 mesh, 200-300 mesh, silica gel H, etc.), Elution conditions: petroleum ether/acetone, chloroform/acetone, chloroform/methanol), Sephadex LH-20 (chloroform/methanol = 1:1, methanol), MCI resin (methanol/water), preparative TLC and preparative HPLC Equivalent column chromatography was used to separate and identify 14 compounds, 9 of which were known abieta-type diterpenes, namely 6-oxo-abieta-8,11,13-triene-11,12-diol(CS -23,12mg), Taxodone (CS-24,26mg), 11,12,14-trihydroxyabieta-8,11,13-trien-7-one (CS-25,29mg), 6α-hydroxydemethyl cryptojaponol (CS-26 ,38mg), 6,7-dioxo-abieta-8,11,13-triene-11,12-diol (CS-27,15mg), hypargenin C (CS-28,7mg), 6,7-dehydroroyleanone (CS -29, 10mg), 14-deoxycoleon U (CS-30, 12mg), 7,7"-bistaxodione (CS-3136mg); 5 are neoabietic diterpenes, which are two B-ring cleavage products Taxodascens E (CS-32, 28mg) and F (CS-33, 4mg), and 3 normal abietane-type diterpene monomers Taxodascens G (CS-34, 21mg), H (CS-35, 11mg), I ( CS-36, 7 mg), J (CS-37, 11 mg).

The characteristics of each compound are as follows:

6-oxo-abieta-8,11,13-triene-11,12-diol (23)

Pale yellow powder, MF: C ₂₀ H ₂₈ O ₃ , EI-MS m/z 316 (100%, M ⁺ ), ¹ H-NMR (CDCl ₃ , 400MHz): δ6.40 (1H, s), 5.93( 1H,br),5.02(1H,br),3.70(1H,d,J=20.0Hz),3.37(1H,d,J=20.0Hz),3.24-3.16(1H,m),3.03(1H,sept ,J=6.8Hz),2.65(1H,s),1.81-1.53(5H,m),1.35(3H,s),1.25(3H,s),1.23(3H,d,J=6.8Hz),1.08 (3H,d,J=6.8Hz), 1.02(3H,s).

Taxodone(24)

Orange-red amorphous powder, MF: C ₂₀ H ₂₈ O ₃ , EI-MS m/z: 316 (26%, M ⁺ ), ¹ H-NMR (CDCl ₃ , 400MHz): δ1.11 (3H, d, J=6.8Hz),1.13(3H,d,J=6.8Hz),1.13(3H,s),1.19(3H,s),1.19(3H,s),1.40(2H,m),1.43(2H, m,),1.54(1H,m),1.55(2H,m),1.63(2H,m),1.65(2H,m),2.89(2H,m),3.04(1H,Sept,J＝6.8Hz) , 4.67 (1H, br d, J = 10.0Hz), 6.52 (1H, d, J = 2.7Hz), 6.79 (1H, s), 7.46 (1H, s).

11,12,14-trihydroxyabieta-8,11,13-trien-7-one (25)

Yellow amorphous powder, MF:C ₂₀ H ₂₈ O ₄ , HR-EI-MS m/z 332.1996[M] ⁺ , ¹ H-NMR(CDCl ₃ ):δ12.16(1H,br s),6.33(1H br s), 4.83(1H, br s), 3.42(1H, sept, J=6.8Hz), 2.98(1H, brd, J=12.4Hz), 2.57(1H, m), 1.78(1H, overlap), 1.42-1.72(2H,m),1.24-1.44(2H,m),1.53(1H,m),1.32(6H,d,J＝6.8Hz),1.34(3H,s),0.94(3H,s) ,0.92(3H,s).

6α-Hydroxydemethylcryptojaponol (26)

Pale yellow solid, MF: C ₂₀ H ₂₈ O ₄ , EIMS m/z: 332[M] ⁺ (58), 317(25), 303(100), 299(17), 285(7), 271(6 ), 247(46), 235(29), 231(13), 219(23); ¹ H NMR (500MHz, CDCl ₃ ): δ1.19(3H,s), 1.24(3H,s), 1.27and 1.29(each 3H,d,J＝6.9Hz)7Hz),1.30and 1.51(each 1H,m),1.54(3H,s),1.53(1H,m),1.72(1H,m),1.82(1H, d,J=13Hz), 2.97(1H,m),3.00(1H,sept,J=6.9Hz),3.87(1H,d,J=2.5Hz),4.59(1H,dd,J=13,2.5Hz ), 5.64 and 5.68 (each 1H, br s), 7.58 (1H, s).

6,7-dioxo-abieta-8,11,13-triene-11,12-diol (27)

Pale yellow solid, MF: C ₂₀ H ₂₆ O ₄ , EI MS: m/z 330 (100%, M ⁺ ), ¹ H-NMR (CDCl ₃ , 400MHz): δ7.63 (1H, s), 6.60( 1H,s),3.14-2.84(2H,m),2.71(1H,s),1.2-1.8(5H,m),1.41(3H,s),1.27(3H,s),1.08(3H,d, J=6.5Hz), 1.07(3H,d, J=6.5Hz), 1.05(3H,s).

Hypargenin C(28)

White amorphous powder, MF: C ₂₀ H ₂₆ O ₃ .EI MS: m/z 314 (50%, M ⁺ ), ¹ H NMR (CDCl ₃ , 400MHz): δ1.12 (3H, s), 1.12 ( 3H,s),1.32(3H,s),1.18(3H,d,J=7Hz),1.20(3H,d,J=7Hz),1.0-1.8(m),2.60(1H,s),2.8( 1H,dd,J=3,12Hz), 3.08(1H,m), 6.24(1H,s), 6.90(1H,s), 7.58(1H,s).

6,7-Dehydroroyleanone (29)

Red needle crystal, mp: 171℃, MF: C ₂₀ H ₂₆ O ₃ ; EI MS: m/z 314 (79%, M ⁺ ), ¹ H-NMR (CDCl ₃ , 400MHz): δ0.98 (3H ,s),1.01(3H,s),1.03(3H,s),1.21(3H,d,J=7.1Hz),1.22(3H,d,J=7.2Hz),1.25(2H,m),1.43 (2H,td,J=13.3Hz),1.51(2H,m),1.62(2H,m),1.71(2H,m),2.14(1H,t,J=3.1Hz),2.89(2H,d, J = 3.4Hz), 3.17 (1H, hept, J = 7.1Hz), 6.46 (1H, dd, J = 3.0Hz), 6.81 (1H, dd, J = 3.1Hz), 7.34 (1H, s).

14-Deoxycoleon U(30)

Pale yellow powder, MF: C ₂₀ H ₂₆ O ₄ , EI MS: m/z 330[M] ⁺ (49), 315(11), 287(13), 274(11), 260(100), 248( 17), 245(16), 233(11), 217(9), ¹ H NMR (500MHz, CDCl ₃ ): δ1.27and1.30(each 3H, d, J＝6.8Hz), 1.41(1H, m ),1.43and 1.44(each 3H,s),1.61and 1.86(each 1H,m),1.71and 2.93(each 1H,m),2.01(1H,td,J＝13and 5Hz),3.04(1H,quint) ,7.09(1H,s),7.70(1H,s).

7,7" - Bistaxodione (31)

Red amorphous powder, MF: C ₄₀ H ₅₀ O ₆ , EI MS: m/z 626 (100%, M ⁺ ), ¹ H-NMR (CDCl3, 400MHz) δ7.63 (2H, s), 6.60 (2H ,d,J＝1.2Hz,H-14),2.98(2H,d sept,J＝1.2,7Hz), 2.71(2H,s),1.08,1.10(each 6H,d,J＝7Hz),1.41, 1.24, 1.05 (each 6H, s).

Taxodascens E(32)

White solid, MF: C ₁₉ H ₂₄ O ₄ , ESI-MS: m/z 317[M+H] ⁺ , ¹ H-NMR (CDCl ₃ , 400MHz): δ6.37 (1H,d,J=1.2Hz ),3.16-2.95(1H,m),2.87(1H,dd,J＝13.8,1.6Hz),2.31(1H,s),1.18-1.76(m),1.27(6H,s),1.14(6H, s), 1.12 (3H, d, J = 2.0 Hz).

Taxodascens F(33)

White solid, MF: C ₂₁ H ₃₀ O ₅ , ESI-MS: m/z 363[M+H] ⁺ , ¹ H-NMR (CDCl ₃ , 400MHz): δ7.70(1H,s), 6.39(1H ,d,J=1.3Hz),4.31(1H,d,J=18.2Hz),4.07(1H,d,J=18.2Hz),3.61(1H,s),3.46(3H,s),3.03-2.94 (1H,m),2.55(1H,d,J=14.1Hz),1.07-1.90(m),1.22(3H,s),1.16(3H,d,J=2.8Hz),1.15(3H,d, J=2.7Hz), 0.83(3H,s), 0.81(3H,s).

Taxodascens G(34)

Light yellow amorphous powder, MF: C ₂₆ H ₃₈ O ₆ , ESI-MS: m/z 447[M+H] ⁺ , ¹ H-NMR(CDCl ₃ , 400MHz): δ7.11(1H,s), 5.20(1H,dd,J=12.0,3.3Hz),4.48(1H,d,J=3.3Hz),3.88(1H,dt,J=8.7,5.8Hz),3.59(1H,dt,J=8.7, 5.8Hz), 3.16(1H,p,J＝7.1Hz), 2.64(1H,d,J＝12.6Hz), 2.15(3H,s), 0.89-1.70(m), 1.36(3H,s), 1.22 (3H,d,J=7.2Hz), 1.19(3H,d,J=7.1Hz), 1.17(3H,s), 0.95(3H,s).

Taxodascens H(35)

Light yellow amorphous powder, MF: C ₂₃ H ₃₄ O ₄ , ESI-MS: m/z 375[M+H] ⁺ , ¹ H-NMR(CDCl ₃ , 400MHz): δ6.97(1H,s), 5.68(1H,s),5.06(1H,d,J=6.6Hz),4.52(1H,dd,J=8.9,6.6Hz),3.04(1H,p,J=6.9Hz),2.84(1H,d ,J=12.2Hz),1.30-1.85(m),1.54(1H,d,J=8.9Hz),1.47(3H,s),1.43(3H,s),1.27(3H,d,J=6.8Hz ), 1.25(3H,d,J=6.8Hz), 1.24(3H,s), 1.13(3H,s), 1.08(3H,s).

Taxodascens I(36)

Light yellow amorphous powder, MF: C ₂₄ H ₃₆ O ₄ , ESI-MS: m/z 389[M+H] ⁺ , ¹ H-NMR(CDCl ₃ , 400MHz): δ6.88(1H,s), 4.56(1H,d,J=4.8Hz),3.91(1H,d,J=4.6Hz),3.88(1H,m),3.59(1H,m),2.980-2.86(1H,m),2.58(1H ,d,J=12.8Hz),2.30(1H,s),1.04-1.82(m),1.62(3H,s),1.56(3H,s),1.12(3H,d,J=4.0Hz),1.10 (3H,d,J=4.0Hz), 1.00(3H,s), 0.99(3H,t).

Taxodascens J(37)

Yellow amorphous powder, MF:C ₄₀ H ₅₆ O ₆ , MS(ESI)m/z 632(M) ⁺ ,1H-NMR(CDCl ₃ ,400MHz):δ7.09(1H,s),6.86(1H, s), 4.64 (1H, d, J = 4.5Hz), 4.58 (1H, d, J = 8.7Hz), 4.46 (1H, dd, J = 12.4, 4.5Hz), 3.94 (1H, dd, J = 11.3 ,8.8Hz),2.97-3.09(2H,m),2.82-2.93(2H,m),2.26(1H,d,J＝12.4Hz),1.74(1H,d,J＝11.1Hz),1.15-1.80 (m),1.52(3H,s),1.42(3H,s),1.40(3H,s),1.31(3H,s),1.24-1.28(15H,m),1.13(3H,s);

1.4. Separation of abietane-type diterpenoids from Chinese fir bark

Chinese fir (Cunninghamia lanceolata) bark (13.5Kg) was crushed into coarse powder, soaked in industrial acetone at room temperature three times, each time for 7 days, combined extracts, concentrated under reduced pressure to remove acetone. Add 10L of 35°C distilled water to dissolve, extract three times with petroleum ether, dichloromethane, and ethyl acetate respectively, and concentrate under reduced pressure to obtain 15g of petroleum ether, 160g of dichloromethane, and 470g of ethyl acetate. The dichloromethane part is passed through normal phase silica gel (100-200 mesh, 200-300 mesh, silica gel H, etc.), elution conditions: petroleum ether/acetone, chloroform/acetone, chloroform/methanol), Sephadex LH-20 (chloroform/methanol =1:1, methanol), MCI resin (methanol/water, ethanol/water), preparative thin-layer and preparative high-performance liquid phase column chromatography for separation, a total of 24 compounds were separated and identified, 14 of which were new compounds, respectively Bicunningine A (27,25 mg), Bicunningine B (28,5mg), Bicunningine C (29,70mg), Bicunningine D (30,6mg), Bicunningine E (31,4mg ), Bicunningine F(32,14mg), Bicunningine G(33,34mg), Bicunningine H(34,25mg), Bicunningine I(35,25mg), Bicunningine J(36,26mg), Bicunningine K(37,15mg), Bicunningine L (38,5mg), 2 abietane diterpene monomers Cunningine A (43,20mg), Cunningine B (44,9mg); 2 known compounds were identified as: Hinokiol (42) and 6α-hydroxy- 7-oxo-ferruginol (52, same as 6).

The above compounds have the following physicochemical and spectroscopic characteristics:

Bicunningine A(27)

White amorphous powder. MF: C ₄₀ H ₅₀ O ₄ , ESIMS: m/z 595.4[M+H] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ2.06and 2.51(m, each 1H, H-1), 2.54and 3.03( m,each 1H,H-2),2.15(d,J=7.4Hz,1H,H-5),5.29(dd,J=7.4,8.1Hz,1H,H-6),4.91(d,J= 8.1Hz,1H,H-7),6.63(s,1H,H-11),6.55(s,1H,H-14),2.88(sept,J＝6.9Hz,1H,H-15),0.69and 0.87(d,J＝6.9Hz,each 3H,H-16,17),1.35,1.42,1.55(s,each 3H,H-18,19,20); 2.67(m,2H,H-1") ,3.12and 3.37(m,each1H,H-2"),2.52(sept,J＝6.9Hz,1H,H-4"),7.17(d,J＝8.1Hz,1H,H-6"),7.61 (d,J=8.1Hz,1H,H-7"),7.51(s,1H,H-14"),3.05(sept,J=6.9Hz,1H,H-15"),1.27(d,J ＝6.9Hz,6H,H-16",17"),1.03and 1.04(d,J＝6.9Hz,each 3H,H-18",19"),2.50(s,3H,H-20").

Bicunningine B(28)

White amorphous powder. MF:C ₄₀ H ₅₂ O ₄ ,ESIMS:m/z 595.4[MH] ⁺ . ¹ H NMR(600MHz,CDCl ₃ )δ1.79and 2.19(m,each 1H,H-1),1.93and 3.11(m, each 1H,H-2),3.41(m,1H,H-3),1.76(d,J=8.3Hz,1H,H-5),5.23(dd,J=7.8,8.3Hz,1H,H- 6), 4.76(d, J=7.8Hz, 1H, H-7), 6.63(s, 1H, H-11), 6.50(s, 1H, H-14), 2.89(sept, J=6.9Hz, 1H,H-15),0.69and 0.86(d,J＝6.9Hz,each 3H,H-16,17),1.17,1.28,1.30(s,each 3H,H-18,19,20); 2.66( m,2H,H-1"),3.11and 3.35(m,each 1H,H-2"),2.51(sept,J＝6.9Hz,1H,H-4"),7.15(d,J＝8.2Hz ,1H,H-6"),7.60(d,J=8.2Hz,1H,H-7"),7.50(s,1H,H-14"),3.11(sept,J=6.9Hz,1H,H -15"), 1.27and1.28(d, J＝6.9Hz, each 3H, H-16", 17"), 1.01and 1.03(d, J＝6.9Hz, each 3H, H-18", 19"),2.48(s,3H,H-20").

Bicunningine C(29)

Light yellow amorphous powder. _{( _ _} ^_ _{_ _} ^{_ _} _{_} m,each1H,H-1),1.90(m,2H,H-2),3.33(m,1H,H-3),1.66(d,J=7.9Hz,1H,H-5),4.93(d ,J=7.9Hz,1H,H-6),3.98(s,1H,H-7),6.73(s,1H,H-11),6.88(s,1H,H-14),3.14(sept, J＝6.8Hz, H-15), 1.20and 1.24(d, J＝6.8Hz, each 3H, H-16,17), 1.01, 1.15and 1.36(s, each 3H, H-18,19,20) ,3.13(s,3H,7-OMe);1.87and 2.25(m,each 1H,H-1"),1.90(m,2H,H-2"),3.41(m,1H,H-3") ,2.16(dd,J＝3.0,3.0Hz,1H,H-5"),5.89(dd,J＝9.7,3.0Hz,1H,H-6"),6.56(dd,J＝9.7,3.0Hz, 1H,H-7"),6.98(s,1H,H-11"),6.94(s,1H,H-14"),3.12(sept,J＝6.9Hz,1H,H-15"),1.03 and 1.04(d,J＝6.9Hz,each 3H,H-16",17"),1.11,1.05and 1.12(s,each 3H,H-18",19",20").

Bicunningine D(30)

Light yellow amorphous powder. MF:C ₄₁ H ₅₈ O ₄ ,EIMS:m/z 614[M] ⁺ , ¹ H NMR(600MHz,CDCl ₃ )δ1.30and 1.48(m,each 1H,H-1),1.67and 1.83(m, each 1H,H-2),1.57and 2.14(m,each 1H,H-3),1.65(d,J＝8.4Hz,1H,H-5),4.89(d,J＝8.4Hz,1H,H -6),3.96(s,1H,H-7),6.75(s,1H,H-11),6.88(s,1H,H-14),3.14(sept,J＝6.9Hz,H-15) ,1.20and 1.24(d,J＝6.9Hz,each 3H,H-16,17),0.92,1.16and 1.39(s,each 3H,H-18,19,20),3.16(s,3H,7- OMe); 1.85and 2.17(m, each 1H, H-1"), 1.84and 1.93(m, each 1H, H-2"), 3.41(m, 1H, H-3"), 2.17(m, 1H ,H-5"),5.89(d,J＝9.7Hz,1H,H-6"),6.55(d,J＝9.7Hz,1H,H-7"),7.01(s,1H,H-11 "),6.95(s,1H,H-14"),3.14(sept,J＝6.9Hz,1H,H-15"),1.04and 1.07(d,J＝6.9Hz, each 3H,H-16",17"),1.11,1.07 and 1.12(s,each 3H,H-18",19",20").

Bicunningine E(31)

Light yellow amorphous powder. MF: C ₄₁ H ₅₆ O ₆ , ESIMS: m/z 645[M+H] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ2.09and 2.13(m, each 1H, H-1), 1.98and 2.16( m,each 1H,H-2),3.30(m,1H,H-3),1.66(m,1H,H-5),5.04(d,J＝8.1Hz,1H,H-6),4.13( s,1H,H-7),6.89(s,1H,H-11),7.03(s,1H,H-14),3.28(sept,J=6.9Hz,H-15),1.15and 1.20(d ,J=6.9Hz,each 3H,H-16,17),0.92,1.10and 1.40(s,each 3H,H-18,19,20),3.20(s,3H,7-OMe);1.74and 2.58 (m,each 1H,H-1"),1.85(m,2H,H-2"),3.43(m,1H,H-3"),6.42(s,1H,H-6"),7.36( s,1H,H-11"),7.92(s,1H,H-14"),3.19(sept,J＝6.9Hz,1H,H-15"),1.02and 1.14(d,J＝6.9Hz, each 3H,H-16",17"),1.33,1.37 and 1.66(s,each 3H,H-18",19",20").

Bicunningine F(32)

Light yellow amorphous powder. MF: C ₄₁ H ₅₈ O ₅ , ESIMS: m/z 629[M+H] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ1.46and 1.71(m, each 1H, H-1), 1.73and 2.52( m,each 1H,H-2),3.35(m,1H,H-3),1.70(d,J＝8.2Hz,1H,H-5),4.92(d,J＝8.2Hz,1H,H- 6),4.02(s,1H,H-7),6.75(s,1H,H-11),6.90(s,1H,H-14),3.16(sept,J＝6.9Hz,1H,H-15 ), 1.19 and 1.23 (d, J＝6.9Hz, each 3H, H-16, 17), 0.96, 1.13 and 1.40 (s, each 3H, H-18, 19, 20), 3.15 (s, 3H, 7 -OMe); 1.74and 2.16(m,each 1H,H-1"),1.13and 1.78(m,each 1H,H-2"),1.38and 1.88(m,each 1H,H-3"),6.49 (s,1H,H-6"),7.25(s,1H,H-11"),8.02(s,1H,H-14"),3.16(sept,J＝6.9Hz,1H,H-15" ),1.05and 1.14(d,J＝6.9Hz,each 3H,H-16",17"),1.28,1.38and 1.64(s,each 3H,H-18",19",20").

Bicunningine G(33)

Light yellow amorphous powder. MF:C ₄₁ H ₅₆ O ₅ ,ESIMS:m/z 629[M+H] ⁺ . ¹ H NMR(600MHz,CDCl ₃ )δ1.58and 2.12(m,each 1H,H-1),1.68(m, 2H,H-2),1.30and 1.46(m,each 1H,H-3),1.70(d,J＝8.2Hz,1H,H-5),4.92(d,J＝8.2Hz,1H,H- 6), 4.02(s, 1H, H-7), 6.77(s, 1H, H-11), 6.89(s, 1H, H-14), 3.18(sept, J=6.9Hz, 1H, H-15 ), 1.19 and 1.22 (d, J＝6.9Hz, each 3H, H-16, 17), 0.87, 1.13 and 1.39 (s, each 3H, H-18, 19, 20), 3.14 (s, 3H, 7 -OMe); 1.78and 2.52(m, each 1H, H-1"), 1.41and 2.02(m, each 1H, H-2"), 3.45(dd, J＝11.3, 4.7Hz, 1H, H-3 "),6.56(s,1H,H-6"),7.27(s,1H,H-11"),8.01(s,1H,H-14"),3.20(sept,J＝6.9Hz,1H, H-15"),1.06and1.14(d,J＝6.9Hz,each 3H,H-16",17"),1.33,1.37and 1.64(s,each 3H,H-18",19",20 ").

Bicunningine H(34)

Light yellow amorphous powder. MF: C ₄₀ H ₅₂ O ₅ , ESIMS: m/z 623[M+H] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ2.54and 2.86(m, each 1H, H-1), 2.09and 2.49( m,each 1H,H-2),1.77(s,1H,H-5),4.96-4.97(m,2H,H-6,7),6.65(s,1H,H-11),6.95(s ,1H,H-14),2.69(sept,J=6.9Hz,1H,H-15),0.89and 1.21(d,J=6.9Hz,each 3H,H-16,17),1.03and 1.47(s ,each 3H,H-18,19),3.08and 4.40(d,J＝8.4Hz,2H,H-20);1.84and 2.30(m,each 1H,H-1"),1.83and 1.95(m, each 1H,H-2"),3.42(dd,J＝10.8,4.3Hz,1H,H-3"),2.14(d,J＝3.0Hz,1H,H-5"),5.90(d,J ＝9.6,3.0Hz,1H,H-6"),6.53(d,J＝9.6,3.0Hz,1H,H-7"),6.89(s,1H,H-11"),6.86(s,1H ,H-14"),3.19(sept,J=6.9Hz,1H,H-15"),0.89and 1.15(d,J=6.9Hz,each 3H,H-16",17"),1.05,1.10 and 1.12(s,each 3H,H-18",19",20").

Bicunningine I(35)

Yellow amorphous powder. MF: C ₄₀ H ₅₀ O ₅ , ESIMS: m/z 611.5[M+H] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ2.50and 2.90(m, each 1H, H-1), 2.17and 2.53( m,each 1H,H-2),1.82(d,J=4.3Hz,1H,H-5),5.16(dd,J=4.3,4.0Hz,1H,H-6),5.19(d,J= 4.0Hz, 1H, H-7), 6.67(s, 1H, H-11), 6.88(s, 1H, H-14), 3.14(sept, J=6.9Hz, 1H, H-15), 1.09and 1.16(d,J＝6.9Hz, each 3H,H-16,17), 1.13and 1.53(s, each 3H,H-18,19), 3.09and 4.46(d,J＝8.6Hz, each 1H,H -20); 2.81(m,2H,H-1"),3.36-3.46(m,2H,H-2"),2.60(sept,J＝6.9Hz,1H,H-4"),7.17and 7.53 (d,J=8.3Hz,each 1H,H-6",7"),7.55(s,1H,H-11"),7.50(s,1H,H-14"),3.09(sept,J= 6.9Hz, 1H, H-15"), 0.90and 1.01(d, J＝6.9Hz, each 3H, H-16", 17"), 1.11and 1.08(d, J＝6.9Hz, each3H, H-18 ",19"),2.50(s,3H.H-20").

Bicunningine J(36)

Yellow amorphous powder. MF: C ₄₀ H ₅₂ O ₄ , ESIMS: m/z 595.4[MH] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ2.78(m,2H,H-1), 3.34(m,2H,H- 2), 2.63(sept, J=6.9Hz, 1H, H-4), 6.79(s, 1H, H-7), 7.22(s, 1H, H-11), 7.36(s, 1H, H-14 ), 3.31(sept, J=6.9Hz, 1H, H-15), 1.29and 1.30(d, J=6.9Hz, each 3H, H-16, 17), 1.13(d, J=6.9Hz, 6H, H-18,19); 2.42(s,3H,H-20); 1.60(dt,J=12.6,4.7Hz,1H,H-1a"),1.91(dt,J=12.6,2.9Hz,1H, H-1b"),1.77(m,2H,H-2"),3.36(m,1H,H-3"),2.15(d,J＝2.7Hz,1H,H-5"),5.95(dd ,J＝9.7,2.7Hz,H-6"),6.63(d,J＝9.7,2.7Hz,1H,H-7"),6.64(s,1H,H-11"),7.07(s,1H ,H-14"),3.19(sept,J=6.9Hz,1H,H-15"),1.21and 1.25(d,J=6.9Hz,each 3H,H-16",17"),1.111.02and 1.02(s,each 3H,H-18",19",20").

Bicunningine K(37)

Yellow amorphous powder. MF: C ₄₀ H ₅₂ O ₄ , ESIMS: m/z 615.3[M+H] ⁺ . ¹ H NMR (600MHz, CDCl ₃ ) δ1.71and 1.86(m, each 1H, H-1), 1.73and 1.88( m,each 1H,H-2),3.31(dd,J=10.9,4.3Hz,1H,H-3),1.73(d,J=9.2Hz,1H,H-5),5.04(d,J= 9.2Hz, 1H, H-6), 3.58(s, 1H, H-7), 6.38(s, 1H, H-11), 5.98(s, 1H, H-14), 3.00(sept, J=6.9 Hz,1H,H-15),1.04and 1.06(d,J＝6.9Hz,each 3H,H-16,17),1.07(d,J＝6.9Hz,6H,H-18,19); 1.34( s,3H,H-20);1.84and2.31(m,each 1H,H-1"),1.97and 1.88(m,each 1H,H-2"),3.40(d,J＝10.9,4.3Hz ,1H,H-3"),2.15(d,J=2.7Hz,1H,H-5"),5.93(dd,J=9.6,2.7Hz,H-6"),6.55(d,J=9.6 ,2.7Hz,1H,H-7"),6.83(s,1H,H-11"),6.97(s,1H,H-14"),3.25(sept,J＝6.9Hz,1H,H-15 "),1.13and 1.14(d,J＝6.9Hz,each 3H,H-16",17"),1.101.05and 1.10(s,each 3H,H-18",19",20").

Bicunningine L(38)

Light yellow amorphous powder. MF:C ₄₁ H ₅₆ O ₅ ,EIMS:m/z 628[M] ⁺ , ¹ H NMR(600MHz,CDCl ₃ )δ1.77and 2.17(m,each 1H,H-1),1.78and 1.91(m, each 1H,H-2),3.38(m,1H,H-3),1.73(d,J=7.9Hz,1H,H-5),5.04(d,J=7.9Hz,1H,H-6) ,4.22(s,1H,H-7),6.76(s,1H,H-11),6.88(s,1H,H-14),3.15(sept,J=6.8Hz,H-15),1.16and 1.21(d, J=6.8Hz, each 3H, H-16, 17), 1.01, 1.14 and 1.41(s, each 3H, H-18, 19, 20), 3.13(s, 3H, 7-OMe); 2.73and 3.04(m,each 1H,H-1"),3.38(m,2H,H-2"),2.72(m,1H,H-4"),7.17(d,J＝8.2Hz,1H, H-6"),7.56(d,J＝8.2Hz,1H,H-7"),7.56(s,1H,H-11"),7.61(s,1H,H-14"),3.28(sept ,J=6.9Hz,1H,H-15"),1.07and1.22(d,J=6.9Hz,each 3H,H-16",17"),1.15(s,6H,H-18",19 "),2.49(s,3H,H-20").

Hinokiol(39)

Light yellow powdery solid, easily soluble in chloroform, acetone, methanol and other organic solvents. MW: 302, MF: C ₂₀ H ₃₀ O ₂ . ¹ H NMR (300MHz, CDCl ₃ ) δ0.87(3H, s, H-18), 1.04(3H, s, H-19), 1.15(3H, s,H-20),1.22(3H,d,H-16),1.24(3H,d,H-17),2.23(d,H-5),2.75(1H,m,H-7a),2.86 (1H,dd,J=16.5,5.5Hz,H-7β),3.08(1H,m,H-15),3.27(1H,dd,J=11.6,4.8Hz,H-3),6.61(1H, s,H-11),6.74(1H,s,H-14).

Cunningine A(40)

Pale yellow powdery solid. MF: C ₂₀ H ₂₄ O ₄ , ESIMS: m/z 331.2[M+H] ⁺ , ¹ H NMR (400MHz, CDCl ₃ ) δ7.90(s,1H,H-14),6.69(s,1H, H-11), 4.46(d, J=12.8Hz, 1H, H-6), 3.16(sept, J=6.9Hz, 1H, H-15), 2.47(d, J=12.8Hz, 1H, H- 5),2.80and 2.52(m,each 1H,H-2),2.33and 2.17(m,each 1H,H-1),1.27(d,J＝6.9Hz,6H,H-16,17),1.49 ,1.39,1.30(s,each 3H,H-18,19,20),

Cunningine B(41)

Pale yellow powdery solid. MF: C ₂₀ H ₂₄ O ₄ , ESIMS: m/z 329.2[M+H] ⁺ , ¹ H NMR (400MHz, CDCl ₃ ) δ8.07(s,1H,H-14),6.89(s,1H, H-11), 3.22(sept, J=6.9Hz, 1H, H-15), 2.77(m, 2H, H-2), 2.41and 2.07(m, each 1H, H-1), 1.28(d, J＝6.9Hz,6H,H-16,17),1.28,1.54,1.58(s,each 3H,H-18,19,20).

1.5. Semi-synthesis and separation of carnosic acid derivatives

1.5.1 Purification of Carnosic acid (CA-1)

Carnosic acid crude product 16g (commercially available, 40%) was dissolved in 25ml of dichloromethane, added saturated NaHCO3 solution 100ml, after no bubbles were produced, placed in a separatory funnel, fully shaken, and the dichloromethane layer was separated Finally, the aqueous phase was washed with dichloromethane for three times, and the dichloromethane layer was discarded. Add 2M HCl solution to the aqueous phase to adjust the pH value to acidic, add 25ml of dichloromethane to extract three times, combine the organic phases, dry over anhydrous sodium sulfate, decolorize with activated carbon, and distill under reduced pressure to obtain pure carnosic acid.

1.5.2. Synthesis of Carnosol (CA-2)

Carnosic acid (6.64g, 20mmol) was dissolved in 6ml of acetic acid, after adding a catalytic equivalent of FeCl ₃ (6mg), 2ml of hydrogen peroxide was added at 15°C, the reaction was kept at 15°C and stirred for 2 hours, then 1ml of concentrated hydrochloric acid, reacted overnight. The reaction solution was diluted to 400ml with water, extracted three times with 200ml of dichloromethane, the organic phases were combined, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure to obtain a brown solid, which was subjected to silica gel column chromatography (petroleum ether: ethyl acetate 10: 1) Purification to obtain 4.62 g (1.4 mmol, yield: 70%) of carnosol. The physical and chemical properties of this compound are as follows:

Carnosol (Carnosol, CA-2), white solid, C ₂₀ H ₂₆ O ₄ , ESI-MS: m/z 331.2[M+H] ⁺ , ¹ H NMR (300MHz, CDCl ₃ ) δ0.86( 3H, s, Me-19), 0.90 (3H, s, Me-18), 1.22 (6H, d, J＝7.0Hz, Me-16/17), 1.31 (1H, dd, J＝3.5, 14.0Hz ,H-3α),1.73(1H,q,H-5),1.90(1H,td,H-6β),1.99(1H,dt,H-2β),2.20(1H,m,H-6α), 2.33, 2.39, 2.46 (1H, td, J = 4.3, 14.0Hz, H-1α), 2.90 (1H, br d, H-1β), 3.08 (1H, sept, J = 7.0Hz, H-15), 5.26(1H,br s,Ar-OH),5.37(1H,dd,J＝1.4,4.0Hz,H-7),5.73(1H,br s,Ar-OH),6.64(1H,s,H- 14).

1.5.3. Synthesis of rosmanol (CA-3)

Carnosol (CA-2) (52.1mg, 0.158mmol) was dissolved in 5ml of acetone, and 5% NaHCO ₃ (6ml, 3.57mmol) was added at room temperature, and the reaction was stirred at room temperature for 12h, then distilled under reduced pressure Acetone was removed, and the reaction solution was acidified by adding 2M HCl and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, distilled off under reduced pressure to dissolve, and the residue was purified by silica gel column chromatography (petroleum ether: acetone 4:1) to obtain rosmanol (rosmanol, CA-3 )36mg, yield 70%. ¹ H NMR (400MHz, Acetone-d ₆ ) δ0.93 (3H, s, Me-19), 1.03 (3H, s, Me-18), 1.22, 1.23 (each 3H, d, J＝7.0Hz, Me -16/17),2.00(1H,td,H-1α),2.21(1H,s,H-5),3.20(1H,sept,J＝7.0Hz,H-15),3.21(1H,br d ,H-1β), 4.57(1H,d,J=3.3Hz,H-6),4.74(1H,d,J=3.3Hz,H-7),6.87(1H,s,H-14); EIMS m/z 346[M] ⁺ (100).

By using different bases and referring to the same operation as above, 7-methoxyrosmanol (CA-4) and 7-ethoxyrosmanol (CA-5) can be obtained, and their ¹ H NMR and EIMS data Consistent with literature (J.Nat.Prod.2002, 65, 986-989.).

1.5.4. Synthesis of Carnosic acid γ-lactone (CA-6)

Carnosic acid (CA-1) (62.8mg, 0.2mmol) and 4-dimethylaminopyridine (36.6mg, 0.3mmol) were dissolved in 10ml of dichloromethane, dicyclohexylcarbodiimide (61.8mg, 0.3 mmol). The reaction solution was stirred overnight at room temperature, filtered, dichloromethane was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: acetone 5:1) to obtain 32 mg of carnosic acid γ-lactone (CA-6) , yield 51%. ¹ H NMR (400MHz, CDCl ₃ ) δ1.09 (3H, s, Me-19), 1.16 (3H, s, Me-18), 1.22 and 1.24 (each 3H, d, J＝7.0Hz, Me-16 /17),1.81and 2.22(each 1H,m,H-1),1.71and 2.00(each 1H,m,H-2),1.39and 2.12(each 1H,m,H-3),1.88(1H, m, H-5), 1.6-1.94 (2H, m, H-6), 3.20 (1H, sept, J=7.0Hz, H-15), 2.57 (2H, m, H-7), 6.66 (1H ,s,H-14); ESIMS m/z 315.2[M+H] ⁺ .

1.5.5.6, Synthesis of 7-dehydrocarnosic acid dimethyl ether (CA-7)

Carnosol (16.5 mg, 0.05 mmol) was dissolved in 2 ml of acetone, and dimethyl sulfate (CH ₃ ) ₂ SO ₄ (5 ul, 0.11 mmol) was added dropwise to the reaction solution. After the dropwise addition, the reaction solution was reacted overnight at room temperature. The reaction solution was distilled off under reduced pressure to remove the organic solvent, and the residue was subjected to gel Sephadex LH-20 (chloroform-methanol 1:1) column chromatography to obtain 17 mg of 6,7-dehydrocarnosic acid dimethyl ether (CA-7a). The rate is 94%. ¹ H NMR (400MHz, CDCl ₃ ) δ6.(1H,d,J=1.3Hz),4.65(1H,dd,J=3.2,0.9),3.88(1H,d,J=3.0Hz),3.69(3H ,s),3.22(1H,m),3.07(1H,s),2.93(1H,m),1.98(1H,s),1.00-2.20(m),1.13(3H,d,J＝6.9Hz) ,1.12(3H,d,J=6.9Hz),1.01(3H,s),0.90(3H,s); EIMS m/z 358(M+).

By controlling the reaction equivalent of dimethyl sulfate and replacing different reaction substrates, phenolic hydroxyl etherification products such as CA-12 and CA-19 can be obtained.

Example 2 Discovery of abietane-type diterpenoids' hypolipidemic activity

The compounds LSP-6-1～LSP-6-6 (final concentration: 5 μM) obtained from the aforementioned separations were used to treat HepG2 cells starved for 12 hours in defatted serum for 24 hours, and 20 μg/ml of fluorescently labeled low-density lipoprotein (DiI-LDL) was added , and incubated at 37°C for 4 hours, gently washed the cells with phosphate buffered saline (PBS) for 5 times, extracted the lipid with isopropanol, and measured the fluorescence readings on a microplate reader (excitation light: 520nm; emission light: 570nm). Then the cells were lysed with 0.2M sodium hydroxide, the protein content was measured, and the value of fluorescence/protein was calculated. The experimental results are shown in Table 1 and Figure 1. At a concentration of 5 μM, the six compounds can significantly increase the uptake of low-density lipoprotein LDL by hepatocyte HepG2.

Table 1. The effect of compounds LSP-6-1～LSP-6-6 on the uptake of low-density lipoprotein in hepatocytes

Compound number LDL uptake rate DMSO 1 ZBM30 (Nagilactone B(7)) 1.46 ^& LSP-6-1 1.11 LSP-6-2 1.26 ^* LSP-6-3 1.16 ^* LSP-6-4 1.29 ^* LSP-6-5 1.26 ^* LSP-6-6 1.33 ^*

(*p<0.05, ^& p<0.01vs control)

HepG2 cells starved for 12 hours in defatted serum were treated with different concentrations of compound LSP-6-6 (concentrations were 5, 10, and 20 μM) for 24 hours, and the uptake rate of low-density lipoprotein by the cells was calculated according to the above method, and the LSP-6 -6 Relationship between dose and activity. The experimental results are shown in Table 2 and Figure 2.

Table 2. The effect of compound LSP-6-6 on the uptake of low-density lipoprotein in hepatocytes at different concentrations

Compound number LDL uptake rate DMSO 1 ZBM30 1.49 ^& LSP-6-6 (5μM) 1.28 ^* LSP-6-6 (10μM) 1.38 ^* LSP-6-6 (20μM) 1.47 ^&

( ^* p<0.05, ^& p<0.01vs control)

The above results indicated that compound LSP-6-6 increased the phagocytosis of LDL by HepG2 cells in a dose-dependent manner; the effect of increasing LDL uptake at a concentration of 20 μM was comparable to that of ZBM30.

Example 3 compound LSP-6-6 up-regulates the expression level of low-density lipoprotein receptor (LDLR) in hepatocytes

Compounds LSP-6-6, berberine (BBR) and ZBM30 were treated with different concentrations of hepatocytes for 24 hours, and after the cells were lysed in the same manner as in Example 2, total cellular RNA was extracted with TRIzol reagent, and 3 μg of RNA was used for M-MLV (reverse transcriptase) was reverse-transcribed into cDNA, and then real-time PCR (real-time quantitative PCR) was performed with LDLR-specific primers, and GAPDH was used as an internal reference gene to compare the regulatory effect of the compound on the LDLR gene. See Figure 3 for the experimental results. The above results showed that the compound LSP-6-6 significantly increased the expression of LDLR gene, increased the expression level of low-density lipoprotein receptor (LDLR) in hepatocytes, and the protein level of LDLR increased with the increase of drug concentration. In terms of the amount of LDLR increase, the activity of LSP-6-6 is equivalent to that of berberine at the same dose, but the dose-effect relationship is more obvious.

Example 4 Test of LDL uptake-promoting activity of abietane-type diterpenoids

The abietane-type diterpenoids (final concentration: 5 μM) isolated from the aforementioned plants or semi-synthesized HepG2 cells were treated with defatted serum starved for 12 hours for 24 hours, and fluorescently labeled low-density lipoprotein (DiI-LDL) was added at 20 μg/ ml, incubated at 37°C for 4 hours, gently washed the cells with phosphate buffered saline (PBS) for 5 times, extracted the lipids with isopropanol, and measured the fluorescence readings on a microplate reader (excitation light: 520nm; emission light: 570nm). Then the cells were lysed with 0.2M sodium hydroxide, the protein content was measured, and the value of fluorescence/protein was calculated. The experimental results of some compounds are shown in Table 3.

Table 3: Effects of some aranane-type diterpenoids on the uptake of low-density lipoprotein in hepatic cells

Compound number LDL uptake rate/DMSO ZBM30 ^& +++ BBR ^& +++ LS-42 ^& + LS-44 ^& +++ LS-53 ^& ++ LS-67 ^& ++ LS-75 ^& ++ CS-6 ^# + CS-8 ^# ++ CS-16 ^& ++ CS-26 _& +

CS-27 ^& + CS-31 ^& + CS-37 ^& ++ CA-1 ^& +++ CA-2 ^# +++

^# p<0.01 ^& p<0.05; +: LDL uptake rate/DMSO 1.20-1.30; ++LDL uptake rate/DMSO 1.31-1.40; +++LDL uptake rate/DMSO 1.40- 1.60

The compound LS-44, CA-1\2, at the same concentration (5uM), the activity of promoting LDL uptake by hepatocytes is equivalent to that of ZBM30.

Example 5 Compound LSP-6-6 orally administered in vivo lipid-lowering efficacy research

1.4.1. Experimental animals

Male Syrian golden hamsters, weighing 100±10 g, were purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences.

1.4.2. Drug preparation

Compound LSP-6-6 was formulated with 5% DMSO, 2% Tween80 and 93% physiological saline to prepare a 15 mg/ml suspension drug solution, which was ultrasonically mixed before use; each animal was given an administration volume of 200 μl/100 g, and given The dosage is 30mg/kg.

1.4.3. Experimental method

After one week of adaptive feeding, the golden hamsters were randomly and evenly divided into groups according to their initial blood lipid and body weight levels. The high-fat model group and the treatment group were given high-fat feed, the treatment group was given 30 mg/kg compound LSP-6-6 by intraperitoneal injection, the high-fat model group was given the same volume of solvent orally, and the normal feed control group was given normal feed. At the beginning of the experiment, blood was collected from the suborbital vein once a week after fasting for 16 hours, and blood lipid indexes were measured. Three weeks later, the golden hamsters were killed by neck dislocation, and the livers were weighed and stored at -80°C until use.

Blood lipid measurement: put the blood at 4°C for two hours to make it stratified, centrifuge at 3000rpm for 15 minutes, draw the upper serum, dilute it 5 times with normal saline, and measure the blood lipid index with an automatic biochemical analyzer.

Statistical method: At least 6 animals in each group were used for statistical analysis with Prism software. ANOVA analysis was used to compare the differences between multiple groups; the t test was used to compare between two groups.

Experimental results

As shown in Figure 4, after the golden hamster fed with high fat was given LSP-6-6 treatment (30mg/kg), compared with the high fat model group, in golden hamster blood indexes TC (serum total cholesterol), TG ( triglycerides), LDL levels decreased significantly.

Through calculation, it was found that compared with the high-fat model group, the reduction percentages of LSP-6-6 on the blood lipids of golden hamsters were:

TC(%) TG(%) LDL-C (%) LSP-6-6 administration 10d 58.37 62.93 36.13 LSP-6-6 administration 20d 75.01 72.80 57.88

After 20 days of LSP-6-6 administration, the reduction rates of blood lipid TC, TG, and LDL levels in the tested animals were close to or exceeded 60%, and LSP-6-6 increased the uptake of LDL by hepatocytes in vitro, which has a significant dose-dependent relationship. , so it is speculated that reducing the dose of LSP-6-6 can also reduce the levels of TC, TG, and LDL in the blood of the tested animals.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (10)

1. A compound shown in the following formula I, its dimer, or the purposes of a pharmaceutically acceptable salt thereof: in, Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl; X is selected from the following group: CHRe, NRe, O, C=Re, C-Re, N; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl; M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl; or Re, Rf are each independently a substituent selected from the following group: Or Re and Rf jointly constitute a structure selected from the following group: -O-, -C ₁ -C ₆ alkylene -, -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene -OC ₁ -C ₆ alkylene -, Or one of Re, Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; Wherein, the substitution means that one or more hydrogen atoms on the group are replaced by =O; Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl; Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or Unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group: halogen, -OH, -COOH; Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-; Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH; Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or form a 5-9 membered epoxy ring or lactone ring together with Rc; R" is selected from the group consisting of H, -COOH, -OH, -Me; Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group; Dotted lines are single or double bonds; and It is a substituted or unsubstituted aromatic ring (benzene ring or benzoquinone); It is characterized in that the compound is used for one or more purposes selected from the following group: a) preparing a pharmaceutical composition for lowering blood fat; b) preparing a pharmaceutical composition for lowering low-density lipoprotein (LDL) levels; c) preparing a pharmaceutical composition for stabilizing LDLR gene stability; d) preparing a pharmaceutical composition for up-regulating LDLR gene expression; e) preparing a pharmaceutical composition for increasing the number of LDLR receptors on the surface of liver cells; f) preparing a pharmaceutical composition for reducing degradation of LDLR receptors; g) preparing a pharmaceutical composition for reducing the concentration of TC and/or TG in blood; h) preparing a pharmaceutical composition for increasing the concentration of high-density lipoprotein (HDL) in blood; i) preparing a pharmaceutical composition for improving liver function damage. 2. purposes as claimed in claim 1, is characterized in that, the dimer of described formula I compound has the structure shown in following formula Ia: In the formula, Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl; The definitions of all the other groups are as described in claim 1. 3. purposes as claimed in claim 1, is characterized in that, X is selected from the following group: CHRe, O, C=Re, C-Re; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ - C ₆ alkyl, -OC ₁ -C ₆ acyl; M is selected from the following group: CHRf, O, C=Rf, C-Rf; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ - C ₆ alkyl, -OC ₁ -C ₆ acyl; Or Re and Rf jointly constitute a structure selected from the following group: -O-, -C ₁ -C ₆ alkylene -, -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene -OC ₁ -C ₆ alkylene -; Or one of Re and Rf together with Ri constitutes a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; wherein, the Substitution means that one or more hydrogen atoms on the group are replaced by =O; Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl; Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted Or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution refers to one of the groups or multiple hydrogen atoms are substituted by substituents selected from the following group: halogen, -OH, -COOH; Ri is selected from the group consisting of H, -COOH, -Me; or Ra and Ri jointly form -C(O)-O-; The definitions of all the other groups are as described in claim 1. 4. The use according to any one of claims 1-3, characterized in that, Ra, Rb, Rc, Rd are each independently selected from the following group: H, -OH, -OMe; X is selected from the following group: CHRe, O, C=Re, C-Re; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, =O; M is selected from the following group: CHRf, O, C=Rf, C-Rf; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, =O; And X and M are not O at the same time; Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₃ alkyl, -OH; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by Substituents selected from the group consisting of: halogen, -OH, -COOH; Rj is H. 5. purposes as claimed in claim 1, is characterized in that, described pharmaceutical composition is also used for being selected from the purposes of following group: j) (in vitro non-therapeutic) increasing the uptake rate of LDL by hepatocytes; k) Up-regulation (non-therapeutically) of the number of LDLR receptors on the surface of hepatocytes in vitro. 6. The use according to claim 1, characterized in that, in the pharmaceutical composition, the effective dose of the compound of formula I is 0.1-50 mg/kg body weight, preferably 1-20 mg/kg body weight. 7. A medicine box, characterized in that, said medicine box contains: (i) a first container, and the active ingredient (a) compound of formula I contained in the first container; or the medicine containing the active ingredient (a); (ii) a second container, and the active ingredient (b) statin drug, or a pharmaceutically acceptable salt thereof; or a drug containing the active ingredient (b) contained in the second container; and (iii) an instruction sheet describing an explanation for reducing the low-density lipoprotein content in the body of a user by co-administering the active ingredient (a) and the active ingredient (b). 8. A compound represented by the following formula I, or a dimer thereof: in, Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl; X is selected from the group consisting of CHRe, NRe, O, C=Re, C-Re, N; M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, said Re, Rf are each independently a substituent selected from the following group: Or Re and Rf jointly constitute a structure selected from the group consisting of: -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene - OC ₁ -C ₆ alkylene-, Or one of Re, Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; Wherein, the substitution means that one or more hydrogen atoms on the group are replaced by =O; Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl; Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or Unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group: halogen, -OH, -COOH; Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-; Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH; Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or together with Rc form a 5-9 membered oxygen-containing ring (preferably an epoxy ring or a lactone ring); R" is selected from the group consisting of H, -COOH, -OH, -Me; Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group; Dotted lines are single or double bonds; and It is a substituted or unsubstituted aromatic ring (benzene ring or quinone). 9. A compound represented by the following formula I, or a dimer thereof: in, Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl; X is O; M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl; Or said Rf is a substituent selected from the following group: Or Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; wherein, the Substitution means that one or more hydrogen atoms on the group are replaced by =O; Rg and Rh are each independently selected from the following group: H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or Unsubstituted C ₁ -C ₆ alkoxy; or Rg and Rh jointly form =O; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: halogen, -OH, -COOH; Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-; Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH; Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or together with Rc form a 5-9 membered oxygen-containing ring (preferably an epoxy ring or a lactone ring); R" is selected from the group consisting of H, -COOH, -OH, -Me; Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group; Dotted lines are single or double bonds; and It is a substituted or unsubstituted aromatic ring (benzene ring or quinone). 10. A compound represented by the following formula I, or a dimer thereof: in, Ra, Rb, Rc, Rd are each independently selected from the following group: H, halogen, -OH, -OMe, O, -OC ₁ -C ₆ alkyl, -CHO, -OC ₁ -C ₆ acyl; X is selected from the following group: CHRe, NRe, O, C=Re, C-Re, N; wherein, the Re is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl; M is selected from the following group: CHRf, NRf, O, C=Rf, C-Rf, N; wherein, the Rf is selected from the following group: H, -CHO, -COOH, -OH, -OMe, =O, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ acyl; Or described Re, Rf are each independently a substituent selected from the following group: Or Re and Rf jointly constitute a structure selected from the group consisting of: -C ₁ -C ₆ alkylene -O-, -OC ₁ -C ₆ alkylene -O-, or -C ₁ -C ₆ alkylene - OC ₁ -C ₆ alkylene-, Or one of Re, Rf and one of Ri or Rg together constitute a substituted or unsubstituted -C ₁ -C ₆ alkylene-, a substituted or unsubstituted -C ₁ -C ₆ alkylene-O-; Wherein, the substitution means that one or more hydrogen atoms on the group are replaced by =O; Or the chemical bond between X, M and together constitutes a group selected from the following group: CH-OR', wherein R' is selected from the following group: H, C ₁ -C ₆ alkyl; Rg is selected from the group consisting of H, -COOH, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, substituted or unsubstituted C ₁ -C ₆ alkoxy, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or unsubstituted C ₁ -C ₆ alkoxy; wherein, the substitution means that one or more hydrogen atoms on the group are substituted by a substituent selected from the following group: halogen, -OH, -COOH; Rh is selected from the group consisting of -COOH, -OH, substituted or unsubstituted C ₁ -C ₆ alkylene-OH, substituted or unsubstituted C ₁ -C ₆ alkylene-COOH, -COO-substituted or unsubstituted Substituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, -COO-substituted or unsubstituted C ₁ -C ₆ alkoxy; wherein, the substitution means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group: halogen, -OH, -COOH; Ri is selected from the group consisting of H, -COOH, -OH, -Me; or Ra and Ri together form -C(O)-O-; Rj is one or more substituents on ring A, and said Rj are each independently selected from the following group: H, =O, substituted or unsubstituted C ₁ -C ₆ alkyl, -OH, -COOH, -COO-substituted or unsubstituted C ₁ -C ₆ alkyl, -COO-substituted or unsubstituted C ₆ -C ₁₀ aryl, -COO-substituted or unsubstituted C ₁ -C ₁₀ heteroaryl, wherein , the substitution refers to being substituted by one or more substituents selected from the following group: halogen, -OH, -COOH; Rk is selected from the following group: H, halogen, C1-C6 alkyl, -CH ₂ OH, -CH ₂ OCOOR, -COOR; or together with Rc form a 5-9 membered oxygen-containing ring (preferably an epoxy ring or a lactone ring); R" is selected from the group consisting of H, -COOH, -OH, -Me; Unless otherwise specified, the "substitution" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following group: carboxyl, C ₁ -C ₆ alkyl, phenyl, C ₃ -C ₆ ring Alkyl, C ₁ -C ₁₀ ester group, halogen, C ₁ -C ₁₀ alkyl-oxyl group, C ₂ -C ₁₀ acyl group, hydroxyl, hydroxy-C ₁ -C ₁₀ alkylene group; Dotted lines are single or double bonds; and It is a substituted or unsubstituted aromatic ring (benzene ring or quinone).